Ki-67 Protein Expression Research Articles

Trichosanthin cooperates with Granzyme B to restrain tumor formation in tongue squamous cell carcinoma

BackgroundTongue squamous cell carcinoma (TSCC) is a common type of oral cancer, with a relatively poor prognosis and low post-treatment survival rate. Various strategies and novel drugs to treat TSCC are emerging and under investigation. Trichosanthin (TCS), extracted from the root tubers of Tian-Hua-Fen, has been found to have multiple biological and pharmacological functions, including inhibiting the growth of cancer cells. Granzyme B (GrzB) is a common toxic protein secreted by natural killer cells and cytotoxic T cells. Our group has reported that TCS combined with GrzB might be a superior approach to inhibit liver tumor progression, but data relating to the use of this combination to treat TSCC remain limited. The aim of this study was to examine the effectiveness of TCS on TSCC processes and underlying mechanisms.MethodsFirst, we screened the potential antitumor activity of TCS using two types of SCC cell lines. Subsequently, a subcutaneous squamous cell carcinoma xenograft model in nude mice was established. These model mice were randomly divided into four groups and treated as follows: control group, TCS treatment group, GrzB treatment group, and TCS/GrzB combination treatment group. Various tumorigenesis parameters, such as Ki67, PCNA, caspase-3, Bcl-2 and VEGFA, et al., were performed to determine the effects of these treatments on tumor development.ResultsScreening confirmed that the SCC25 line exhibited greater sensitivity than the SCC15 line to TCS in vitro studies. TCS or GrzB treatment significantly inhibited tumor growth compared with the inhibition seen in the control group. The TCS/GrzB combination inhibited tumor growth more than either drug alone. TCS treatment inhibited tumor proliferation by downregulating Ki67 and Bcl2 protein expression while accelerating tumor apoptosis. In the TCS/GrzB-treated group, expression of Ki67 was further downregulated, while the level of activated caspase-3 was increased, compared with their expression in either of the single drug treatment groups.ConclusionThese results suggest that the TCS/GrzB combination could represent an effective immunotherapy for TSCC.

The Pathology of Morphine-Inhibited Nerve Repair and Morphine-Induced Nerve Damage Is Mediated via Endoplasmic Reticulum Stress.

ObjectiveThe aim of the present study was to observe the pathological damage in the cerebral cortex of rats under acute morphine exposure (AME) and different durations of morphine dependence (MD), explore whether endoplasmic reticulum stress (ERS) is involved in the damage process, and assess the effect of morphine exposure on the proliferation and differentiation of newborn neurons.MethodsRat models of AME and different durations of MD were established. Pathological changes in cortical neurons were assessed by hematoxylin and eosin (H&E) and thionine staining. The expression of nuclear receptor-related factor 1 (NURR1) and that of the ERS-related proteins glucose-regulated protein 78 (GRP78), p-eIF2α, activating transcription factor 6 (ATF6), and CHOP in cortical neurons was assessed by immunohistochemistry. Double immunofluorescence labeling was used to observe the expression of Ki-67.ResultsH&E and thionine staining revealed that AME resulted in pyknotic changes in cortical neurons. With prolonged morphine exposure, the number of pyknotic neurons was significantly increased, the protein expression of Ki-67 and NURR1 was significantly decreased, and the protein levels of GRP78, p-eIF2α, ATF6, and CHOP showed marked dynamic changes.ConclusionAME and different durations of MD caused varying degrees of pathological changes in the cortex. Furthermore, the dynamic changes observed in ERS-related protein expression suggested that ERS may be associated with cortical injury. Different durations of MD inhibited the proliferation, differentiation, and migration of newborn neurons, which may affect the nerve repair process after injury.

Abstract GS4-09: Correlative studies of the breast cancer index (HOXB13/IL17BR) and ER, PR, AR, AR/ER ratio and Ki67 for prediction of extended endocrine benefit: A trans-aTTom study

Abstract Background: Several biomarkers such as estrogen receptor (ER), progesterone receptor (PR), androgen receptor (AR) and Ki67 have been implicated in the pathogenesis and/or as prognostic biomarkers of breast cancer, and are utilized to determine treatment. Given the heterogeneity of response to endocrine therapy, however, predictive biomarkers are critical to better individualize patient care. Previous results from the Trans-aTTom study demonstrated that the Breast Cancer Index HOXB13/IL17BR [BCI (H/I)] biomarker significantly predicts extended endocrine benefit from 10 vs 5y of tamoxifen. In this correlative study, the predictive activity of BCI (H/I) was compared with ER, PR, AR and Ki67 protein expression in node positive patients treated in the aTTom trial. Methods: Patients with available tumor tissue and biomarker analyses were included. ER, PR, AR and Ki67 were centrally assessed by immunohistochemistry (IHC) utilizing tissue microarrays. BCI gene expression analysis by RT-PCR was performed blinded to clinical outcome. Multivariate Cox models adjusting for age, tumor size, tumor grade and HER2 status were used to assess the significance of the interaction between treatment and each biomarker as continuous variables. 17-year risk of recurrence, as a function of each continuous biomarker, was estimated from Cox models in each of the 2 treatment arms. Results: Analysis of 789 HR+, N+ patients showed a weak negative correlation between BCI (H/I) and ER, PR, and AR expression whereas Ki67 and the AR/ER ratio showed no correlation (ER, cor=−0.18; PR, cor=−0.25; AR, cor=−0.14; Ki67, cor=0.04; AR/ER ratio, cor=0.02). The interaction between BCI (H/I) and extended tamoxifen treatment was significant (p=0.014). In addition, analysis of risk of recurrence as a function of continuous BCI (H/I) demonstrated that the magnitude in the reduction in recurrence risk with extended tamoxifen correlated with increasing H/I levels. In contrast, interaction P values were nonsignificant (ER, p=0.829; PR, p=0.659; AR, p=0.783; Ki67, p=0.865; AR/ER ratio, p=0.835) and the magnitude of endocrine benefit did not correlate with expression levels of any of other biomarkers. Conclusion: Results from this post-hoc analysis of the Trans-aTTom study demonstrated that whereas BCI(H/I) is a significant predictive biomarker of endocrine response, analysis of ER, PR, AR, Ki67 and AR/ER expression showed no interaction with treatment, and lacked the ability to predict benefit of extended tamoxifen in HR+ early stage breast cancer. These results add to the growing body of evidence that BCI (H/I) is distinct in its ability to predict benefit from therapy and interrogates distinct tumor biology that is not captured by other traditional biomarkers. Citation Format: Dennis C Sgroi, Kai Treuner, Yi Zhang, Tammy Piper, Ranelle Salunga, Ikhlaaq Ahmed, Lucy Doos, Sarah Thornber, Karen J Taylor, Elena F Brachtel, Sarah Pirrie, Catherine A Schnabel, Daniel W Rea, John MS Bartlett. Correlative studies of the breast cancer index (HOXB13/IL17BR) and ER, PR, AR, AR/ER ratio and Ki67 for prediction of extended endocrine benefit: A trans-aTTom study [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr GS4-09.

Quantitative perfusion histogram parameters of dynamic contrast-enhanced MRI to identify different pathological types of uterine leiomyoma.

:To explore the value of quantitative perfusion histogram parameters of dynamic contrast-enhanced magnetic resonance imaging （DCE-MRI） in pathological classification of uterine leiomyoma and its correlation with Ki-67 protein expression. Thirty five patients with uterine leiomyoma confirmed by operation and pathology at Shaoxing People's Hospital from October 2015 to September 2017 were analyzed retrospectively，including 15 cases of ordinary type，8 cases of cellular type and 12 cases of degenerative type. All patients were examined by pelvic DCE-MRI before operation，and the histogram parameters （median，mean，skewness，kurtosis，energy，entropy） of various quantitative perfusion parameters，including volume transport constant （K），rate constant （K），extravascular extracellular space distribute volume per unit tissue volume （V），blood plasma volume per unit volume of tissue （V） were calculated，and the efficacy of different parameters in pathological classification of uterine leiomyoma was evaluated by ROC curve. The expression of Ki-67 protein in uterine leiomyoma was detected by immunohistochemical method，and the correlation between histogram parameters and Ki-67 protein expression was analyzed by Pearson and Spearman correlation analysis. The median and mean values of K，K，V and V in the cellular group were higher than those in the degenerative group and the ordinary group（<0.05 or <0.01），while the skewness of V，the skewness and kurtosis of K in the cellular group were lower than those in the ordinary group （all <0.05）. The entropy of K in the cellular group was higher than that in the degenerative group and the ordinary group （all < 0.05）. The entropy of V in the cellular group was higher than that in the ordinary group （<0.01）. The median，mean，skewness of K，median and mean of K，median and mean of V，median，mean，energy and entropy of V were correlated with Ki-67 expression（all <0.05）. The results of ROC curve analysis showed that the median threshold of K was 0.994/min，the sensitivity and specificity for the diagnosis of cellular uterine leiomyoma were 100.0% and 77.8% respectively，and the area under the ROC curve was 0.949. When the mean threshold of K was 1.170/min，the sensitivity and specificity for diagnosing cellular uterine leiomyoma were 100.0% and 77.8% respectively，and the area under the ROC curve was 0.958. The area under the ROC curve of K （entropy），K （median，mean），V （median，mean，entropy） in the diagnosis of cellular uterine leiomyoma were 0.755-0.907. :DCE-MRI quantitative perfusion histogram parameters have high diagnostic value in differentiating pathological types of uterine leiomyoma，especially for cellular uterine leiomyoma.

The prognostic relevance of p53 and Ki-67 to chemotherapy sensitivity and prognosis in triple-negative breast cancer.

BackgroundThis study aimed to explore the prognostic function of p53 and Ki-67 protein expression in chemotherapy sensitivity and prognosis in triple-negative breast cancer (TNBC).MethodsPatients who were confirmed with TNBC in Wenzhou Geriatric Hospital and Wenzhou Hospital of Traditional Chinese Medicine (including the Oncology Department, Tumor Surgery Department, and Gynecology Department) between January 2006 and February 2018 were included in this study. The expression of p53 and Ki-67 detected by immunohistochemistry, the rate of recurrence, and the objective curative effect evaluation at the end of the first-line rescue treatment were recorded for all patients.ResultsThe patients were followed up to August 2020, and the median follow-up time was 9 years and 4 months. A total of 285 patients with TNBC were enrolled in the study. The patients ranged in age from 19 to 76 years old, with an average age of 53 years. The overall recurrence rate among the patients was 31.58%. The majority of cases (68.07%) were pathological stage I. The overall positive expression rates of Ki-67 and p53 were 53.33% and 56.84%, respectively. In the TNBC recurrence group, the positive rates of p53 and Ki-67 were 71.11% and 82.22%, respectively, which were significantly higher than those in the non-recurrence group. The positive rates of p53 and Ki-67 in the chemosensitive group were 96.05% and 92.11%, respectively, which were significantly higher than those in the non-chemosensitive group. Among all the TNBC patients, 128 patients had positive expression of both p53 and Ki-67, and 101 patients had negative expression of both p53 and Ki-67. The chemosensitivity rate of TNBC patients with positive expression of both Ki-67 and p53 was 98.53%, and that of TNBC patients with negative expression of both Ki-67 and p53 was 0.00%. The difference was statistically significant. The recurrence rate in TNBC patients with positive expression of both Ki-67 and p53 was 53.13%, and that in patients with negative expression of both Ki-67 and p53 was 6.93%. The difference was statistically significant.ConclusionsThe expression of p53 and Ki-67 had prognostic relevance to chemotherapy sensitivity and prognosis in TNBC patients.

FSH Receptor and Ki-67 Protein Expressions of Endometrial Macaque in Mid-Luteal Phase after Exposure to Various Doses of Recombinant FSH and Fixed-Dose of GnRH Agonist

FSH Receptor and Ki-67 Protein Expressions of Endometrial Macaque in Mid-Luteal Phase after Exposure to Various Doses of Recombinant FSH and Fixed-Dose of GnRH Agonist

Allanblackia floribunda Seed Extract Attenuates the Ethanol-Induced Gastric Ulcer in Rats via the Inhibition of TNF-α and INF-γ Levels and Modulation in the Expression of Ki67 Protein.

Allanblackia floribunda has been used to treat an upset stomach in African traditional medicine, but its efficacy and safety have not been scientifically studied. The present research is aimed at assessing the antiulcer property of the seed extract of the plant to validate its traditional claim. Rats were pretreated with three doses of aqueous extract of A. floribunda (AFE) at 30, 100, and 300 mg/kg or omeprazole 10 mg/kg for 1 hr before the acute gastric ulcer was induced by oral administration of 5 mL/kg of 98% ethanol. The animals were sacrificed under anesthesia, and the stomach and blood were collected. The gross histology of the stomach, percentage protection conferred by the treatment, gastric pH, and serum TNF-α and INF-γ were assessed as well as the expression of Ki67 antigens. The antioxidant properties as well as the acute toxicity profile of the plant extract were also assessed. The results show that A. floribunda conferred significant protection on the rats against gastric ulceration with % protection of 46.15, 57.69, and 65.38 for AFE 30, 100, and 300 mg/kg, respectively, as well as 69.23% for omeprazole 10 mg/kg. The plant extract caused marked reductions in gastric pH, TNF-α, and INF-γ with statistical significance (p < 0.001) for AFE 300 mg/kg and omeprazole 10 mg/kg. Also, the plant showed good antioxidant activity comparable to gallic acid. Furthermore, the plant extract modulated the expression of Ki67 antigens. All animals survived the 14-day delayed toxicity test with no significant differences in physical, hematological, and biochemical parameters between rats orally administered with supratherapeutic doses of AFE (5000 mg/kg) or normal saline. The study established that the gastroprotective effect of the seed extract of A. floribunda is attributable to its antisecretory, antioxidant, and anti-inflammatory properties. Additionally, the plant was found to promote ulcer healing via the modulation of the expression Ki67 and was safe at supratherapeutic doses.

Capsaicin Inhibits Proliferation and Induces Apoptosis in Breast Cancer by Down-Regulating FBI-1-Mediated NF-κB Pathway.

BackgroundAs a natural compound extracted from a variety of hot peppers, capsaicin has drawn increasing attention to its anti-cancer effects against multiple human cancers including breast cancer. FBI-1 is a major proto-oncogene negatively regulating the transcription of many tumor suppressor genes, and plays a vital role in tumorigenesis and progression. However, whether FBI-1 is involved in capsaicin-induced breast cancer suppression has yet to be ascertained. This study aimed to investigate the effects of capsaicin on proliferation and apoptosis and its association with FBI-1 expression in breast cancer.MethodsCCK-8 and morphological observation assay were employed to detect cell proliferation. Flow cytometry and TUNEL assay were conducted to detect cell apoptosis. RNA interference technique was used to overexpress or silence FBI-1 expression. qRT-PCR and/or Western blot analysis were applied to detect the protein expression of FBI-1, Ki-67, Bcl-2, Bax, cleaved-Caspase 3, Survivin and NF-κB p65. Xenograft model in nude mice was established to assess the in vivo effects.ResultsCapsaicin significantly inhibited proliferation and induced apoptosis in breast cancer in vitro and in vivo, along with decreased FBI-1, Ki-67, Bcl-2 and Survivin protein expression, increased Bax protein expression and activated Caspase 3. Furthermore, FBI-1 overexpression obviously attenuated the capsaicin-induced anti-proliferation and pro-apoptosis effect, accompanied with the above-mentioned proteins reversed, whereas FBI-1 silencing generated exactly the opposite response. In addition, as a target gene of FBI-1, NF-κB was inactivated by p65 nuclear translocation suppressed with capsaicin treatment, which was perceptibly weakened with FBI-1 overexpression or enhanced with FBI-1 silencing.ConclusionThis study reveals that FBI-1 is closely involved in capsaicin-induced anti-proliferation and pro-apoptosis of breast cancer. The underlying mechanism may be related to down-regulation of FBI-1-mediated NF-κB pathway. Targeting FBI-1 with capsaicin may be a promising therapeutic strategy in patients with breast cancer.

Maternal Gestational &lt;i&gt;Bifidobacterium bifidum&lt;/i&gt; TMC3115 Treatment Shapes Construction of Offspring Gut Microbiota and Development of Immune System and Induces Immune Tolerance to Food Allergens

Background: Our previous studies has proved that Bifidobacterium bifidum TMC3115 (TMC3115) treatment during infancy could protect neonates from adulthood allergy. However, whether it could still protect offspring from allergic diseases when treatment during maternal pregnancy and the underlying mechanisms remain unclear. Methods: Pregnant BALB/c mice were gavaged with TMC3115 until delivery. Offspring were reared until weaning (postnatal day 21, PND21) and then sensitized with ovalbumin (OVA) until PND49. Vaginal, gut, and breast milk microbiota were sequenced. Immunoglobulin and cytokines, splenic and colonic gene expressions, and fecal short chain fatty acids (SCFAs) were measured. Findings: After maternal TMC3115 treatment, the microbiota of maternal breast milk from newborn offspring and gut microbiota of neonates and at weaning were changed the most, and levels of serum and breast milk immunoglobulin and of splenic cytokines as well as mRNA expression of colonic intestinal development indicators were all significantly altered in offspring in different stages. Synchronous changes in the specific microbiota were found in the maternal gut, vagina, and breast milk and in the offspring gut microbiota, such as Proteobacteria. After sensitization with OVA, there was no significant change in levels of serum IgE or OVA-specific IgE/IgG1 in the offspring in TMC3115 group; however, IgM, expression of colonic Ki67, Muc2, and tight junction proteins and production of fecal SCFAs were significantly increased as were the relative abundances of Lactobacillus and Lachnospiraceae NK4A136 group. Interpretation: Our results suggested that the day of birth and weaning were the important “window of opportunity” for neonatal gut microbiota construction and that maternal treatment with TMC3115 could play a profound modulatory effect on the development of the immune system in offspring at different stages. Maternal TMC3115 treatment may induce immune tolerance to allergens in OVA-stimulated offspring and enhance intestinal tissue development and barrier function and primary immune response to allergens through modulating gut microbiota and SCFA production. Funding Information: This work was supported by the National Natural Science Foundation of China (Grant number: 81372982). Declaration of Interests: The authors declare that they have no conflict of interest. Ethics Approval Statement: The animal experiment facility and animals used for the present study were officially approved by the Experimental Animal Management Committee of Sichuan Government. Experimental protocols were approved by the West China School of Public Health Medical Ethics Committee of Sichuan University (Approved number: SYXK2018-011).

Commentary: Expression of p53 and Ki-67 proteins in patients with increasing severity and duration of pterygium.

Commentary: Expression of p53 and Ki-67 proteins in patients with increasing severity and duration of pterygium.

Investigation of the Inhibitory Effect of Platycodin D in Human Transitional Cell Carcinoma Cell Line 5637

Platycodin D is an active component isolated from Chinese herb Platycodonis radix with various pharmacological activities, such as antitussive, expectorant, anti-inflammatory, and analgesic effects. Interestingly, platycodin D also exerts anticancer effects against several types of cancer. However, few studies on the anti-tumour effects of platycodin against urinary bladder cancer have been reported. In this study, we explored the anti-tumour effect of platycodin D against human bladder cancer and its mechanisms in vitro and in vivo. We found that platycodin D had significant anti-proliferative effects on four types of cancer cells, especially the 5637 bladder cancer cell line, and exerted these effects by preventing cell cycle progression from G0/G1 to S phase, down-regulating Ki-67 and cyclin D1 protein expression and up-regulating P21 protein expression. Furthermore, platycodin D inhibited 5637 cell migration by decreasing twist-related protein 1 (Twist1) and matrix metallopeptidase 2 (MMP2) expression and exerted significant tumour-suppressive effects in tumour-bearing nude mice. Platycodin D also increased caspase-9, caspase-8, caspase-3, and p53 expression and decreased Bcl-2 expression in tumour tissues. Taken together, our results provide a theoretical basis for application of platycodin D in treating urinary bladder cancer.

Liver Kinase B1 (LKB1) Regulates Proliferation and Apoptosis of Non-Small Cell Lung Cancer A549 Cells via Targeting ERK Signaling Pathway

ObjectiveTo study the effect and potential mechanism of LKB1 on non-small cell lung cancer (NSCLC) A549 cells.Material and MethodsA549 cells were divided into control group, LKB1 negative control (NC) group, LKB1 group, ERK inhibitor group and LKB1 + ERK activator group. Cell proliferation and apoptosis were detected by cell counting kit (CCK-8) assay and flow cytometry, respectively. Transwell assay was used to analyze the invasion ability of A549 cells. The expression of apoptosis and ERK signaling pathway-related proteins were studied by Western blot. Furthermore, a nude mouse xenograft model was constructed and treated with LKB1, ERK inhibitor and activator, respectively. The tumor volume and tumor weight were measured. Immunohistochemistry was used to test the expression of Ki-67 protein in tumor tissues, and TUNEL staining was used to test the apoptosis. Moreover, Western blot was used to detect ERK signaling pathway-related proteins in tumor tissues.ResultsCompared with control and NC groups, cell proliferation and invasion were inhibited in ERK inhibitor and LKB1 groups, while apoptosis and apoptosis-related proteins were increased (p < 0.05). Further study showed that ERK activator can reverse the effect of LKB1 in A549 cells. In nude mice, ERK inhibitor and LKB1 can reduce cell tumorigenicity and inhibit proliferation. Apoptosis was increased by ERK inhibitor and LKB1 treatment. Western blot showed that LKB1 and ERK inhibitor could reduce the protein expression of p-ERK1/2. However, the indicators above were the opposite in the ERK activator group.ConclusionLKB1 overexpression can inhibit proliferation and promote apoptosis of NSCLC A549 cells, and its mechanism may be related to inhibition of the ERK signaling pathway.

Effect of RNA interference targeting neuropilin-2 gene on proliferation and apoptosis of colon cancer cell line HCT-8

Objective: To investigate the effects of down-regulation of expression of neuropilin-2 (NRP-2) by RNA interference (RNAi) technique on proliferation and apoptosis of HCT-8 colon cancer cells. Methods: NRP2-siRNA and negative control (NControl)-siRNA were transferred into HCT-8 colon cancer cells by liposomes (lip2000) as transfection group and negative control group, and phosphate buffered solution (PBS) was added as blank control group. Quantitative reverse transcription PCR (RT-qPCR) and Western blot were used to detect the transfection effect. The proliferation of cells in the three groups was examined by cell counting kit (CCK) assay, colony-forming unit assay and Ki-67 protein staining assay, respectively. Moreover, the apoptosis of cells in the three groups was determined by acridine orange/propranidine iodide (AO/PI) staining method. Results: The results of RT-qPCR and Western blot showed that the relative expression of NRP-2 mRNA and the content of NRP-2 protein in the transfer group decreased (0.46±0.05 vs 0.99±0.05 and 1.00±0.06; 1.04±0.06 vs 1.73±0.09 and 1.65±0.11) (all P<0.05). The results of CCK-8 demonstrated that the optical density of transfection group was significantly lower than that of the negative control group and the blank control group(24 h: 0.53±0.04 vs 0.82±0.07 and 0.87±0.07; 48 h: 0.54±0.05 vs 1.00±0.09 and 1.17±0.05; 72 h: 0.75±0.05 vs 1.31±0.13 and 1.50±0.03; 96 h:1.05±0.04 vs 1.46±0.09 and 1.86±0.06) (all P<0.05). The results of colony-forming unit assay indicated that the proliferation ability of the cells in the transfer group was significantly lower than that in the other two groups (134.67±8.74 vs 245.33±19.14 and 300.33±14.01, P<0.05). The results of Ki-67 protein staining assay showed that compared with the negative control group and blank control group, the expression of Ki-67 protein was significantly decreased in the transfection group (5.93±0.22 vs 8.36±0.09 and 8.70±0.21, P<0.05). The results of AO/PI assay revealed that the ratio of apoptotic cells to living cells in the transfer group was significantly higher than that in the other two groups (0.43±0.07 vs 0.14±0.04 and 0.11±0.04, P<0.05). Conclusion: The proliferation ability of HCT-8 colon cancer cells decreases, and the apoptosis ability increases by decreasing the expression of NRP-2.

Clinical and Subtypes of Breast Cancer in Indonesia

Background: Breast cancer is still a global public health problem, especially in women and is the second leading cause of death after lung cancer. Indonesia is a country with a high incidence of breast among women. Methods: This study used a retrospective study of breast cancer patients registered at Division of Surgical Oncology in Indonesia. Information about demographic, clinical and pathological characteristics of patients for a period of 5 years (2014 to 2020) were collected and described. Results: Information for 1260 cases of breast cancer who underwent IHC examination with an average age of 53.73 years of patients, with the most age above 50 years (63.6%) were collected. On IHC examination, 58.6% positive estrogen receptors were found, positive progesterone receptors were 55.4%, Her2 was positive 37.7% and high Ki67 protein was 56.9%. The most common breast cancer subtypes were luminal B (43.2%), followed by luminal A (21.7%), TNBC (18.1%), and Her2 (17%), with the highest stages being IIIB (31.7%).Conclusions: The breast cancer was presented at higher stage, with more cases with estrogen or progesterone receptors are positive with low frequency of Her2 positivity and high in Ki67 protein expression.&#x0D;

ENDOMETRIAL POLYPS. QUESTIONS OF ETIOLOGY AND PATHOGENESIS

Endometrial polyps are a fairly common type of pathological endometrial hyperplastic process, but the molecular and pathogenetic mechanism underlying their formation and progression remains unclear. However, today their development is explained by a change in the level of normally hormonal reception and secretion of women more often in the premenopausal period or a dysregulation of mitotic activity in endometrial cells. The aim of our study is to study the etiology and pathogenesis of endometrial polyps among women of different age groups using general scientific and morphological research methods. According to histological studies of biopsy samples (scrapings) from the uterine cavity, taken from the archive of the pathological anatomical department of the 442nd military district hospital named after Z.P. Soloviev for the period 2017-2019, revealed pathomorphological features of endometrial polyps among women of different age groups with metrorrhagia and associated pathologies: uterine fibroids, cervical canal polyps (of the same etiology), adenomyosis, infertility. More often, endometrial polyps are combined with uterine myoma (54%) and adenomyosis (43%), which emphasizes the similarity of premorbid background, the same clinical manifestations of diseases and their clinical and pathological features. The main factor in the development of endometrial polyps among women of reproductive age and the premenopausal period in age is a change in hormonal levels, namely, an increase in the secretion and reception of estrogen and a decrease in the reception and secretion of progesterone. And among postmenopausal women undergoing hormonal and antitumor therapy against the background of endometrial atrophy, the formation of endometrial polyps is associated with a change in the regulation of mitotic activity in endometrial cells, which is detected by the expression of Ki-67 protein.

Impact of p53, HIF1a, Ki-67, CA-9, and GLUT1 Expression on Treatment Outcomes in Locally Advanced Cervical Cancer Patients Treated With Definitive Chemoradiation Therapy

The objective of this study was to assess the association between pretreatment p53, hypoxia inducible factor 1a (HIF1a), Ki-67, carbonic anhydrase-9 (CA-9), and glucose transporter 1 (GLUT1) expression in locally advanced cervical cancer patients treated definitively with concurrent chemoradiation therapy (CRT) and treatment outcomes including overall survival (OS), progression-free survival (PFS), local-regional control (LC), and distant metastases-free survival (DMFS). Twenty-eight patients treated definitively and consecutively for cervical cancer with CRT had p53, HIF1a, Ki-67, CA-9, and GLUT1 protein expression assessed and scored semiquantitatively by 3 pathologists, blinded to the treatment outcomes. Outcomes were stratified by p53 (H-score: <15 vs. ≥15), HIF1a (H-score: <95 vs. ≥95), Ki-67 (labeling index <41% vs. ≥41%), CA-9 (H-score: <15 vs. ≥15), and GLUT1 (H-score: <175 vs. ≥175) expression. OS, PFS, LC, and DMFS rates were calculated using the Kaplan-Meier method, and differences between groups were evaluated by the log-rank test. Notable clinical characteristics of the cohort included median age of 51 years (range: 32 to 74 y), FIGO stage IIB disease (57.2%), clinical node-negative disease (64.3%), squamous cell carcinoma (89.3%), and adenocarcinoma (10.7%). Treatment outcomes included 5-year OS (57.2%), PFS (48.1%), LC (72.1%), and DMFS (62.9%). For HIF1a H-score <95 and ≥95, the 5-year OS (52.0% and 68.4%, P=0.58), PFS (53.0% and 40.9%, P=0.75), LC (71.6% and 68.2%, P=0.92), and DMFS (59.7% and 52.0%, P=0.91) were not significantly different. For Ki-67 labeling index <41% and ≥41%, the 5-year OS (44.9% and 66.6%, P=0.35), PFS (38.9% and 55.4%, P=0.53), LC (57.7% and 85.7%, P=0.22), and DMFS (67.3% and 61.0%, P=0.94) were not significantly different. For CA-9 H-score <15 and ≥15, the 5-year OS (54.4% and 66.7%, P=0.39), PFS (57.3% and 40.0%, P=0.87), LC (70.0% and 70.0%, P=0.95), and DMFS (70.0% and 46.7%, P=0.94) were not significantly different. For GLUT1 H-score <175 and ≥175, the 5-year OS (43.6% and 43.6%, P=0.32), PFS (55.6% and 49.5%, P=0.72), LC (72.9% and 71.5%, P=0.97), and DMFS (62.5% and 59.6%, P=0.76) were not significantly different. For p53, H-score <15 and ≥15, the 5-year OS (62% and 53%), PFS (63% and 30.3%), LC (87.5% and 52%), and DMFS (79.6% and 41.6%). In this study population, HIF1a, Ki-67, CA-9, and GLUT1 expression did not predict treatment response or outcomes in locally advanced cervical cancer patients treated definitively with CRT. There was a nonstatistically significant trend towards worse outcomes with p53 expression.

Effect of salvianolic acid A on the proliferation and apoptosis in esophageal cancer cells and the underlying mechanisms.

To explore the effect of salvianolic acid A (SalA) on the proliferation and apoptosis in esophageal cancer cell line KYSE-150 and the possible mechanisms. The esophageal cancer cells were randomly divided into 4 groups: a control group, a 10 μmol/L SalA group, a 25 μmol/L SalA group, and a 50 μmol/L SalA group. Cell counting kit-8 (CCK-8) was used to detect the cell proliferation activity. Flow cytometry was used to detect cell cycle distribution and cell apoptosis rate. Western blotting was used to detect the protein expression of cell proliferation maker Ki-67, cyclin D1, cyclin-dependent kinase 4 (CDK4), CDK6, B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X (Bax), cleaved-caspase-9, cleaved-caspase-3, phosphatidylinositol 3-kinase (PI3K), phosphorylated protein kinase B (p-Akt) and mechanistic target of rapamycin (mTOR). Compared with the control group, the cell proliferation activity was significantly reduced (P<0.01); the cells in the G1 phase were significantly increased, and the S phase cells were significantly reduced (both P<0.01); the cell apoptosis rate was significantly increased (P<0.01) in the SalA groups at different concentration; the expression levels of Ki-67, cyclin D1, CDK4, CDK6, Bcl-2, PI3K, p-Akt and mTOR were decreased significantly, but the expression levels of p21, Bax, cl-caspase-9 and cl-caspase-3 were increased significantly in the SalA groups at different concentration (all P<0.01). SalA can inhibit the proliferation and induce G1 phase arrest and apoptosis in the esophageal cancer cell line KYSE-150, which may be related to the activation of PI3K/Akt/mTOR signal pathway.

5-Azacytidine Depletes Hematopoietic Stem Cells and Synergizes with an Anti-CD117 Antibody to Augment Engraftment of Donor Stem Cells in Immunocompetent Mice

Monoclonal antibody (mAb)-targeting of CD117 (c-Kit) present on hematopoietic stem cells (HSC) is an emerging conditioning strategy for hematopoietic cell transplantation (HCT) to replace standard-of-care alkylator and radiation therapy used to deplete recipient HSC and permit donor HSC engraftment. In preclinical and clinical studies anti-CD117 mAbs, which inhibit stem cell factor (SCF) from binding to CD117, have shown single agent efficacy in permitting donor HSC engraftment in immunocompromised recipients. However, preclinical data suggest that anti-CD117 mAb, ACK2, used as a single agent is insufficient to enable HSC engraftment in immunocompetent mice. Hence, there is a desire to potentiate the effect of naked anti-CD117 mAbs to permit targeted elimination of host HSC for a range of immune competent bone marrow (BM) states, from monogenic disorders, e.g., hemoglobinopathies, to BM clonal malignant diseases. Here, we show that ACK2 synergizes with the widely used hypomethylating agent, 5-Azacytidine (AZA), to deplete HSC and enable robust long-term HSC engraftment in immunocompetent mice. The cell depleting effect of AZA or ACK2+AZA was first tested on mature and primitive hematopoietic cells of C57BL/6 (B6) mice. Mice received AZA at 5mg/kg i.p. for 5 days either alone or combined with ACK2 at 500 µg i.v. 5 days prior to AZA. BM and spleen were harvested days 6, 10 and 20 after the first AZA dose and analyzed by extended immunophenotype. For HCT studies both congenic [B6 (H-2b, Thy1.1, CD45.1) into B6 recipients (H-2b, Thy1.1, CD45.1/CD45.2)] and allogeneic [B6 (H-2b, Thy1.1, CD45.1) into BALB.B recipients (H-2b, Thy1.2, CD45.2)] transplantation mouse models were tested. Single agent AZA induced rapid depletion of CD117+ cells in the BM, including LT-HSC (Lin-Sca+Kit+[LSK]CD150+CD48-) and ST-HSC (LSKCD150-CD48-). In vivo HSC depletion by AZA has not been previously reported, and was surprising given that most HSC are quiescent, and non-proliferating cells are known to be insensitive to the toxicities of AZA. Examination of HSC populations in the spleen did not suggest that mobilization of LT- or ST-HSC to secondary organs caused the decrease in BM. Rather, AZA-induced myelosuppression was followed by rapid expansion of myeloid-biased MPP and increased HSC proliferation as assessed by Ki67 intracellular protein expression. We hypothesized, that the in vivo HSC-depletive effects of AZA might result from increased sensitivity of proliferating HSC to AZA. Hence, the in vitro effects of different AZA concentrations were tested on proliferating HSC. In both mouse and human HSC cultures, supplemented with SCF and TPO, we observed that AZA reduced the number of cells in a dose dependent manner and decreased cell viability at concentrations of ≥ 0.5 µg/ml. In vivo, combined ACK2+AZA led to more robust HSC depletion and prolonged time to HSC recovery compared to AZA alone. We then evaluated if blockade of CD117 with its ligand SCF by ACK2 was the cause of this prolonged HSC-depletion. 2B8 mAb, which also binds mouse CD117, but only partially inhibits CD117/SCF signaling, did not result in augmented in vivo HSC depletion (Figure 1A). We further tested if immunocompetent mice treated with ACK2+AZA could be "rescued" with exogenous recombinant SCF. SCF-treatment resulted in faster recovery of LT-HSC. Together these studies suggest that the synergy of ACK2+AZA on the depth and duration of HSC-depletion depends of the disruption of CD117/SCF binding. Finally, we tested ACK2+AZA conditioning on donor stem cell engraftment in immunocompetent mice. Transplantation of 15-20x106 WBM cells following ACK2+AZA conditioning led to significantly increased stable mean multilineage donor chimerism of &amp;gt;60% in the congenic (Figure 1B) and &amp;gt;20% in the allogeneic setting. Single agent AZA enabled sustained donor engraftment with levels of myeloid donor chimerism of 10-15%, serving as proof of principle that AZA treatment can indeed clear HSC niches and permit engraftment of true HSC. Similar to WBM grafts, 5x104 purified HSC (LSK) engrafted successfully in the ACK2+AZA group, reaching long-term (26 weeks) mean donor myeloid chimerism &amp;gt;25% in both congenic and allogeneic settings. Our studies reveal a previously unknown effect of AZA on BM HSC and provide a platform for the clinical use of a novel less toxic non-myeloablative conditioning strategy for HCT, which is aimed at the rapid translation into clinical practice. Disclosures Pang: Jasper Therapeutics, Inc: Current Employment, Current equity holder in private company. Shizuru:Jasper Therapeutics, Inc: Current equity holder in private company, Membership on an entity's Board of Directors or advisory committees.

Immunohistochemical features of benign endometrial hyperplasia in premenopausal women

Hyperproliferative diseases of the endometrium play an important role in the structure of gynecological pathology, which are a spectrum of irregular morphological changes. Particularly difficult is evaluation of the phenotypic characteristics of the endometrium hyperplastic processes (EHP) in premenopausal women in the presence of an unstable menstrual cycle. Diagnosis and EHP prognosis remains a difficult task given that it can occur as focal or diffuse lesions with various structural and cytological differences.Objective of the study: to evaluate the immunohistochemical features (phenotypic variants) of benign endometrial hyperplasia in premenopausal women.Material and methods. 33 premenopausal women with abnormal uterine bleeding and verified benign endometrial hyperplasia were examined. Expression of the α-receptors for estrogens type 1 (ER1), progesterone receptors and Ki-67 nuclear protein in the endometrium stroma and glands was analyzed. Micromorphometry was performed and the D-score was calculated.Results. Prevalence of comorbid lesions in patients was the combination of endometrial hyperplasia and fibroids (51.4%), cases of abnormal uterine bleedings against submucosal fibroids (13.5%), endometrial polyps (8.1%), combinations of EHP and peritoneal endometriosis (10.8%), adenomyosis and myoma (8.1%), or other combinations of endometrial and myometrial proliferative pathology. D-score for surgery averaged 1.78 ± 0.11 indicating a low risk of malignancy. There were changes after treatment in the quantitative presentation of the studied proteins in stroma and endometrial glands. Thus, before treatment in the glands was determined up to 100% of cells containing ER1 in large quantities, while after treatment their number decreased by an average of 20%. Similar dynamics was observed with progesterone receptors activity.Conclusions. The main prognostic significant phenotypes of endometrial proliferative pathology have been identified. After removing of pathologically chanced endometrium and subsequent treatment with dydrogesterone during 6 months there is prognostically positive decreasing in the ER1 density as well as the Ki-67 protein expression

Morphofunctional and immunohistochemical characteristics of the large omentum in ovarian cancer

The aim of the study was to establish morphofunctional and immunohistochemical characteristics of large omentum in women with ovarian cancer.Material and methods. The large omenta of 48 women with ovarian cancer (low-grade differentiated seropapillary adenocarcinoma of high-grade malignancy) of II stage (n=20) and III stage (n=28) were studied. Histological sections were stained with overview histological and immunohistochemical methods (to reveal ki67, P53, CD34, CD7, CD4, CD8, CD61 proteins expression). Results. In patients, the size of the large omentum was characterized by high individual variability; in the presence of metastasis, the size of the omentum was reduced. Intensive development of blood vessels in the organ was noted, but in the presence of metastases stasis of blood corpuscles, leucocytic infiltration, and moderate edema of connective tissue were observed in the organ’s vessels. Areas of lymphoid tissue, both small lymphatic follicles and diffusely located lymphoid tissue, were revealed in the omentum. In most follicles, reactive centers were not marked, and the number of follicles was reduced in the presence of metastases in the omentum. The analysis of CD34+ cells distribution showed that they were identified both in the tumor and in the areas of the omentum adjacent to the tumor, which indicates a pronounced angiogenesis. An irregular distribution of CD7+ and CD8+ and CD4+ cells was revealed in the tumor tissues, as well as in the surroundings. Simultaneously with the expression of P53 protein, ki67 protein expression is revealed in the significant number of tumor cells (including endothelial cells of tumor blood vessels). The proportion of ki67+ cells in the tumor cell population was 60.1±3.3%. The presence of a large number of ki67+cells in the presence of P53 protein expression in them indicates the aggressiveness of the tumor, as well as a disturbance of apoptosis regulatory mechanisms in the cells. Ki67 expression was low in the omentum areas unaffected by metastases, and it was revealed in the certain areas of connective tissue in fibroblastic programmed differentiation cells. Conclusion. The results obtained indicate significant plasticity and reactivity of great omentum in the presence of tumor process in the body and confirm the important role of great omentum in protective reactions.