Epithelial Cancer Tissues Research Articles

Transcriptomics Analysis of Stromal Breast Cancer Tissues Reveals Up-Regulation of GABA-A Receptor Activity

Breast cancer is the second most prevalent cancer overall among newly diagnosed cases and the most common cancer among women. 90% of cancer deaths are caused by metastasis, which is a factor that precedes local invasion, but very little is understood about the molecular mechanisms of invasion and metastasis. Thus, uncovering the root causes of this illness at the Transcriptomics level may result in a cutting-edge method of treating breast cancer. The total RNA microarray processed data from GEO for breast cancer patients was analyzed to discover hidden differences between epithelial breast cancer tissues (ET), stromal breast cancer tissues (ST), normal control epithelial breast cancer tissue samples (EC), and normal control stromal breast cancer tissue samples (SC) at the Transcriptomics level. Therefore, multiple bioinformatics analyses of the transcriptional profiles of 64 samples—including 28 TEC, 28 SCC, 5 EN, and 5 SN controls—received from the NCBI-Bio project were performed in the current study (PRJNA107497). First, ET versus EC and ST vs SC samples showed significant patterns in exploratory data analysis based on gene expression data using principal component analysis (PCA). Following this, 13512 substantially differentially expressed genes (Fold change (>= 1.5), p.adj value 0.1) between these conditions were found by Welch's T-test differential gene expression analysis. This study identifies the main element that may significantly contribute to the spread of breast cancer from epithelial cells to stromal cells in the mammary glands as the gene like the GBRP. This study was also able to identify the affected biological pathways for both the ST vs. SC samples and the ET vs. EC samples as a result of the up-regulated and down-regulated genes. This most surely provides a crucial hint about the cause of the deadly metastatic cancer problem. Finally, the results presented here provide novel insights on breast cancer metastasis.

Single-cell discovery of the scene and potential immunotherapeutic target in hypopharyngeal tumor environment

Hypopharyngeal carcinoma is a cancer with the worst prognosis. We constructed the first single-cell transcriptome map for hypopharyngeal carcinoma and explored its underlying mechanisms. We systematically studied single-cell transcriptome data of 17,599 cells from hypopharyngeal carcinoma and paracancerous tissues. We identified categories of cells by dimensionality reduction and performed further subgroup analysis. Focusing on the potential mechanism in the cellular communication of hypopharyngeal carcinoma, we predicted ligand-receptor interactions and verified them via immunohistochemical and cellular experiments. In total, seven cell types were identified, including epithelial and myeloid cells. Subsequently, subgroup analysis showed significant tumor heterogeneity. Based on the pathological type of squamous cell carcinoma, we focused on intercellular communication between epithelial cells and various cells. We predicted the crosstalk and inferred the regulatory effect of cellular active ligands on the surface receptor of epithelial cells. From the top potential pairs, we focused on the BMPR2 receptor for further research, as it showed significantly higher expression in epithelial cancer tissue than in adjacent tissue. Further bioinformatics analysis, immunohistochemical staining, and cell experiments also confirmed its cancer-promoting effects. Overall, the single-cell perspective revealed complex crosstalk in hypopharyngeal cancer, in which BMPR2 promotes its proliferation and migration, providing a rationale for further study and treatment of this carcinoma.

Research Progress in the Relationship Between P2X7R and Cervical Cancer.

Cervical cancer is one of the most common and serious tumors in women. Finding new biomarkers and therapeutic targets plays an important role in the diagnosis, prognosis, and treatment of cervical cancer. Purinergic ligand-gated ion channel 7 receptor (P2X7R) is a purine ligand cation channel, activated by adenosine triphosphate (ATP). Studies have shown that P2X7R plays an important role in a variety of diseases and cancers. More and more studies have shown that P2X7R is also closely related to cervical cancer; therefore, the role of P2X7R in the development of cervical cancer deserves further discussion. The expression level of P2X7R in uterine epithelial cancer tissues was lower than that of the corresponding normal tissues. P2X7R plays an important role in the apoptotic process of cervical cancer through various mechanisms of action, and both antagonists and agonists of P2X7R can inhibit the proliferation of cervical cancer cells, while P2X7R is involved in the antitumor effect of Atr-I on cervical cancer cells. This review evaluates the current role of P2X7R in cervical cancer in order to develop more specific therapies for cervical cancer. In conclusion, P2X7R may become a biomarker for cervical cancer screening, and even a new target for clinical treatment of cervical cancer.

Expression analysis and function of mitochondrial genome-encoded microRNAs

MicroRNAs (miRNAs) play a significant role in nuclear and mitochondrial anterograde and retrograde signaling. Most of the miRNAs found inside mitochondria are encoded in the nuclear genome, with a few mitochondrial genome-encoded non-coding RNAs having been reported. In this study, we have identified 13 mitochondrial genome-encoded microRNAs (mitomiRs), which were differentially expressed in breast cancer cell lines (MCF-7, MDA-MB-468 and MDA-MB-231), non-malignant breast epithelial cell line (MCF-10A), and normal and breast cancer tissue specimens. We found that mitochondrial DNA (mtDNA) depletion and inhibition of mitochondrial transcription led to reduced expression of mitomiRs in breast cancer cells. MitomiRs physically interacted with Ago2, an RNA-induced silencing complex (RISC) protein, in the cytoplasm and inside mitochondria. MitomiRs regulate the expression of both nuclear and mitochondrial transcripts in breast cancer cells. We showed that mitomiR-5 targets the PPARGC1A gene and regulates mtDNA copy number in breast cancer cells. MitomiRs identified in the present study may be a promising tool for expression and functional analysis in patients with a defective mitochondrial phenotype, including cancer and metabolic syndromes. This article has an associated First Person interview with the first author of the paper.

Biological Activity of an Epilobium angustifolium L. (Fireweed) Infusion after In Vitro Digestion.

The biological activity of an in vitro digested infusion of Epilobium angustifolium (fireweed) was examined in a model system of intestinal epithelial and colon cancer tissues. The content of selected phenolic compounds in the digested aqueous extract of fireweed was determined using HPLC-ESI-QTOF-MS/MS. Biological activity was examined using the human colon adenocarcinoma cell lines HT-29 and CaCo-2 and the human colon epithelial cell line CCD 841 CoTr. Cytotoxicity was assessed by an MTT assay, a Neutral Red uptake assay, May-Grünwald-Giemsa staining, and a label-free Electric Cell-Substrate Impedance Sensing cytotoxicity assay. The effect of the infusion on the growth of selected intestinal bacteria was also examined. The extract inhibited the growth of intestinal cancer cells HT-29. This effect can be attributed to the activity of quercetin and kaempferol, which were the most abundant phenolic compounds found in the extract after in vitro digestion. The cytotoxicity of the fireweed infusion was dose-dependent. The highest decrease in proliferation (by almost 80%) compared to the control was observed in HT-29 line treated with the extract at a concentration of 250 μg/mL. The fireweed infusion did not affect the growth of beneficial intestinal bacteria, but it did significantly inhibit E. coli. The cytotoxic effect of the fireweed extract indicates that it does not lose its biological activity after in vitro digestion. It can be concluded that the fireweed infusion has the potential to be used as a supporting agent in colon cancer therapy.

Correlation between HPV infection and ovarian epithelial cancer diagnosed by Dynamic Contrast-enhanced magnetic resonance imaging information technology under exponential distribution mathematical model

This research aims to analyze the correlation between human papillomavirus (HPV) infection and ovarian epithelial cancer based on exponential distribution mathematical model, so as to provide an experimental basis for the early diagnosis of ovarian epithelial cancer by magnetic resonance imaging (MRI) in the future. In this study, 124 patients with ovarian epithelial cancer tissues pathologically confirmed in our hospital from March 31, 2017 to February 20, 2019 were selected as the experimental group, and 64 patients with normal ovarian tissues were selected as the control group, and the positive rate of HPV infection was detected by the computer cell test (CCT) system. Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) was constructed and adopted to analyze the scanning images. The correlation between HPV infection and pathological type, clinical stage, tissue differentiation degree, and CA125 in serum was studied by exponential distribution mathematical model. The results showed that the positive rate of HPV infection in the ovarian tissues of the experimental group was significantly higher than that of the control group (P < 0.05). Highly differentiated patients accounted for 25.97%, moderately differentiated patients accounted for 43.81%, and lowly differentiated patients accounted for 30.22%. Patients with clinical stage I-II accounted for 24.72%, patients with stage III accounted for 49.11%, and patients with stage IV accounted for 26.17%. HPV infection was significantly correlated with clinical stage III and moderate tissue differentiation by MRI of patients (P < 0.05), and extremely significantly correlated with clinical stage IV and low tissue differentiation by MRI (P < 0.001). The above findings show that the information technology of dynamic contrast-enhanced magnetic resonance imaging can clearly display the lesion metastasis of ovarian epithelial cancer patients with high soft tissue resolution.

Identification of Distinct Immune Subtypes in Colorectal Cancer Based on the Stromal Compartment.

The tumor environment is of vital importance for the incidence and development of colorectal cancer. Increasing evidence in recent years has elaborated the vital role of the tumor environment in cancer subtype classification and patient prognosis, but a comprehensive understanding of the colorectal tumor environment that is purely dependent on the stromal compartment is lacking. To decipher the tumor environment in colorectal cancer and explore the role of its immune context in cancer classification, we performed a gene expression microarray on the stromal compartment of colorectal cancer and adjacent normal tissues. Through the integrated analysis of our data with public gene expression microarray data of stromal and epithelial colorectal cancer tissues processed through laser capture microdissection, we identified four highly connected gene modules representing the biological features of four tissue compartments by applying a weighted gene coexpression network analysis algorithm and classified colorectal cancers into three immune subtypes by adopting a nearest template prediction algorithm. A systematic analysis of the four identified modules mainly reflected the close interplay between the biological changes of intrinsic and extrinsic characteristics at the initiation of colorectal cancer. Colorectal cancers were stratified into three immune subtypes based on gene templates identified from representative gene modules of the stromal compartment: active immune, active stroma, and mixed type. These immune subtypes differed by the immune cell infiltration pattern, expression of immune checkpoint inhibitors, mutation landscape, extent of mutation burden, extent of copy number burden, prognosis and chemotherapeutic sensitivity. Further analysis indicated that activation of the NF-kB signaling pathway was the major mechanism causing the no immune infiltration milieu in the active stroma subtype and that inhibitors of the NF-kB signaling pathway could be candidate drugs for treating patients with an active stroma. Overall, these results suggest that characterizing colorectal cancer by the tumor environment is of vital importance in predicting patients' clinical outcomes and helping guide precision and personalized treatment.

PD-L1 expression on stromal tumor-infiltrating lymphocytes is a favorable prognostic factor in ovarian serous carcinoma

BackgroundPD-L1 expression levels determined by immunostaining are known to be related to the survival rate and prognosis of patients with various types of cancers, as well as to the therapeutic response to immune checkpoint inhibitors. Recently, the U.S. Food and Drug Administration approved an immune checkpoint inhibitor for the treatment of non-small cell lung cancer along with the clones used for PD-L1 immunostaining to predict the resulting response. In this study, we performed PD-L1 immunostaining of tissue microarrays from ovarian epithelial cancer using SP263, an approved clone, and examined the effect of PD-L1 expression on survival rate and prognosis.MethodsTissue microarrays were constructed from ovarian epithelial cancer tissues of 248 patients and PD-L1 immunostaining was performed using the SP263 clone. PD-L1 expression levels in tumor cells, intraepithelial tumor-infiltrating lymphocytes, and stromal tumor-infiltrating lymphocytes were evaluated, and the effect of PD-L1 expression on survival and prognosis was analyzed.ResultsPD-L1 was detected in tumor cells as well as intraepithelial tumor-infiltrating lymphocytes and stromal tumor-infiltrating lymphocytes. It was most frequently expressed in stromal tumor-infiltrating lymphocytes. The Kaplan-Meier curve analysis and log rank test showed that only high stromal PD-L1 expression was associated with increased overall survival in ovarian serous carcinoma. Multivariate and univariate Cox regression analyses revealed that stromal PD-L1 expression was an independent prognostic factor, especially in ovarian serous carcinoma.ConclusionsPD-L1 immunostaining using SP263 was observed in tumor cells as well as intraepithelial and stromal tumor-infiltrating lymphocytes. PD-L1-expressing stromal tumor-infiltrating lymphocytes were associated with an increased overall survival rate and may serve as a favorable prognostic factor in ovarian cancer, particularly serous carcinoma.

Paip1 Indicated Poor Prognosis in Cervical Cancer and Promoted Cervical Carcinogenesis.

PurposeThis study was aimed to investigate the role of poly(A)-binding protein-interacting protein 1 (Paip1) in cervical carcinogenesis.Materials and MethodsThe expression of Paip1 in normal cervical epithelial tissues and cervical cancer (CC) tissues were detected by immunohistochemistry. In vivo and in vitro assays were performed to validate effect of Paip1 on CC progression.ResultsPaip1 was found to be up-regulated in CC, which was linked with shorter survival. Knockdown of Paip1 inhibited cell growth, induced apoptosis and cell cycle arrest in CC cells, whereas its overexpression reversed these effects. The in vivo tumor model confirmed the pro-tumor role of Paip1 in CC growth.ConclusionAltogether, the investigation demonstrated the clinical significance of Paip1 expression, which prompted that the up-regulated of Paip1 can presumably be a potential prognostic and progression marker for CC.

Potential protective effect of ALDH-1 stromal expression against early recurrence of operable breast cancers.

The emerging cancer stem cell (CSC) model proposes that the stem cell niche plays a major role in the risk of cancer recurrence. Enzymatic activity of aldehydes, including aldehyde dehydrogenase 1 (ALDH-1), has been used as a marker of normal and malignant breast stem cells (BSCs). However, the clinical implications of the expression of stem cell markers in the stroma have not yet been investigated. To determine the relationships of ALDH-1 expression, the currently reliable BSCs marker, with clinical characteristics and survival, we used immunohistochemical staining of tissue microarrays from 180 stroma and epithelial cancer tissues in patients diagnosed with operable early breast cancer (stage 0-III). ALDH-1 expression was observed in 93.4% of the stromal cells and in 37.2% of the epithelial cells, and the expression levels between the two cell types were significantly correlated (P=0.001). The stromal expression of ALDH-1 was not correlated with any clinical factors, whereas epithelial expression was significantly correlated with a negative estrogen-receptor status (P<0.001), high proliferation based on Ki-67 expression (P<0.001), and younger age (P=0.04). After 27.8 months of follow up, negative stromal expression of ALDH-1 was significantly correlated with shorter overall survival (positive, 56.9±3.0 months vs. negative, 30.5±3.0 months; P=0.01). Stromal ALDH-1 expression was not directly correlated with known clinical factors, but its expression may play a protective role against early recurrence. Further observation and large-scale studies are needed to validate the clinical implications of ALDH-1 in breast cancer.

Ovarian fibroma/fibrothecoma with elevated serum CA125 level: A cohort of 66 cases.

Ovarian fibroma/fibrothecoma with elevated serum OC125 antigen (CA125) is rarely encountered in clinical practice, and also easily misdiagnosed as epithelial ovarian carcinoma (EOC). The aim of this study was to investigate the clinicopathological features of ovarian fibroma/fibrothecoma with elevated serum CA125.In total, 580 patients who underwent primary surgery and pathologically diagnosed as ovarian fibroma/fibrothecoma were retrospectively analyzed. The clinicopathological parameters were collected and compared between the patients with elevated serum CA125 (>35 U/mL) and without. The immunoreactivity for CA125 in ovarian fibroma/fibrothecoma and epithelial cancer tissues was detected and compared by immunohistochemistry. Univariate and multivariate analyses were performed to identify factors associated with elevated serum CA125 level. The correlation between the immunoreactivity of CA125 in tissue and serum CA125 level was examined by Pearson correlation analysis.Elevated serum CA125 level (range 36.7–1848 u/m) was found in 66 of 580 (11.3%) ovarian fibroma/fibrothecoma patients. Univariate analysis showed that the elevated serum CA125 level was significantly correlated with tumor diameter ≥10 cm (P < .001), ascites (P < .001), and hydrothorax (P < .001). Multivariate analysis revealed that tumor diameter ≥10 cm and ascites were independently associated factors (P < .001 and < .001 respectively). Immunohistochemical staining showed that the expression of CA125 was negative in all fibroma/fibrothecoma tissues, but positive in all EOC tissues, and the immunoreactivity for CA125 was positively correlated with serum CA125 level in the EOC patients (P = .005).The elevated serum CA125 level in ovarian fibroma/fibrothecoma is nontumor originated and occurs more frequently in those with larger size tumor or Meigs syndrome.

Effects of gold nanorods modified with antiepidermal growth factor receptor monoclonal antibody on laryngeal cancer cells.

We aimed to evaluate the photothermal effects of gold nanorods (GNRs) modified with antiepidermal growth factor receptor (EGFR) monoclonal antibody (mAb) on laryngeal cancer cells. EGFR protein expressions in HEP-2 cells, and normal laryngeal epithelial and laryngeal cancer tissues were detected by western blot. The cytotoxicity against HEP-2 and BEAS-2B cells was tested by MTT assay, and the photothermal effects were assessed by near-infrared (NIR) irradiation. The apoptosis of HEP-2 cells was detected by flow cytometry. EGFR expression in laryngeal cancer tissues was significantly higher than that in normal tissues (P < 0.05). The inhibitory effects of GNRs and anti-EGFR mAb/GNRs on the apoptosis of HEP-2 and BEAS-2B cells were enhanced with increasing dose. AntiEGFR mAb/GNRs were more cytotoxic to HEP-2 cells and less cytotoxic to BEAS-2B cells than GNRs. NIR irradiation inhibited cell proliferation, which was enhanced with rising power, accompanied by continuously dropping survival rate. The apoptosis rate of the anti-EGFR mAb/GNRs group was significantly higher than that of the GNRs group, and the apoptosis rate of the irradiation + antiEGFR mAb/GNRs group also significantly exceeded that of the irradiation + GNRs group (P < 0.05). Anti-EGFR mAb/GNRs promoted HEP-2 cell apoptosis more evidently than GNRs did. Functional modification of GNRs augmented the targeted specificity to cancer cells, biocompatibility, and stability. Anti-EGFR mAb/GNRs have great potential in biomedical fields.

JNK and Yorkie drive tumor progression by generating polyploid giant cells in Drosophila.

Epithelial cancer tissues often possess polyploid giant cells, which are thought to be highly oncogenic. However, the mechanisms by which polyploid giant cells are generated in tumor tissues and how such cells contribute to tumor progression remain elusive. We previously noticed in Drosophila imaginal epithelium that cells mutant for the endocytic gene rab5 exhibit enlarged nuclei. Here we find that mutations in endocytic 'neoplastic tumor-suppressor' genes, such as rab5, vps25, erupted, or avalanche result in generation of polyploid giant cells. Genetic analyses on rab5-defective cells reveal that cooperative activation of JNK and Yorkie generates polyploid giant cells via endoreplication. Mechanistically, Yorkie-mediated upregulation of Diap1 cooperates with JNK to downregulate the G2/M cyclin CycB, thereby inducing endoreplication. Interestingly, malignant tumors induced by Ras activation and cell polarity defect also consist of polyploid giant cells, which are generated by JNK and Yorkie-mediated downregulation of CycB. Strikingly, elimination of polyploid giant cells from such malignant tumors by blocking endoreplication strongly suppressed tumor growth and metastatic behavior. Our observations suggest that JNK and Yorkie, two oncogenic proteins activated in many types of human cancers, cooperatively drive tumor progression by generating oncogenic polyploid giant cells.

The significance of RASSF2A promoter methylation in the early diagnosis of epithelial ovarian cancer

Objective To evaluate the methylation status of RASSF2A in epithelial ovarian cancer (EOC) tissues and plasma, and to explore the correlations in two kinds of samples with clinicopathological characteristics. Methods The frequency of methylation of the RASSF2A gene in tissues and corresponding plasma samples from 60 EOC patients, 20 patients with benign ovarian tumors and 10 patients with normal ovarian tissues were detected respectively. The methylation-specific PCR was used to determine the methylation status. Results The frequency of aberrant methylation of RASSF2A was 58.3 % (35/60) in EOC tissues and 43.3 % (26/60) in corresponding plasma samples respectively, however, such hypermethylation was not detected in the benign ovarian tumors and normal ovarian samples, and there was significant difference between them (P 0.05). Conclusions The promoter methylation of RASSF2A gene is an early and frequent epigenetic event in EOC. Detection of promoter hypermethylation in plasma specimens can help the early diagnosis of EOC. Key words: Ovarian neoplasms; Gene, RASSF2A; Serum; Methylation

Overexpression of CXCL3 can enhance the oncogenic potential of prostate cancer

CXCL3 and its receptor CXCR2 were considered to play particularly important roles in the progression of malignancies. However, the investigations about CXCL3/CXCR2 axis in prostate cancer have been poorly involved. Herein we firstly reported our studies on the expression and biological roles of CXCL3 and CXCR2 in prostate cancer. Expression levels of CXCL3 and CXCR2 in prostate cancer cell lines (PC-3, DU145 and LNCaP), immortalized prostate stromal cell line (WPMY-1) and immortalized prostate epithelial cell line (RWPE-1) were investigated by RT-PCR, ELISA and western blot, whereas expression levels of CXCL3 in a prostate tissue microarray were detected by immunohistochemistry. Cell counting kit-8 and transwell assays were, respectively, utilized to determine the effects of exogenous CXCL3 on the cell proliferation and migration. We further examined whether CXCL3 could regulate the expression of genes correlated with prostate tumorigenesis by RT- PCR. Elevated expression of CXCR2 was detected in DU145, LNCaP and RWPE-1. Moreover, high-level CXCL3 can be secreted by PC-3 and RWPE-1, and CXCL3 protein expression level in tissue microarray is concordant with prostate cancer metastasis. Exogenous CXCL3 does not contribute to proliferation, but has a significant effect on migration of prostate cancer cells and RWPE-1. Finally, our data showed that exogenous CXCL3 can regulate the expression of genes including ERK, TP73, NUMB, BAX and NDRG3. Our findings suggest that CXCL3 and its receptor CXCR2 are overexpressed in prostate cancer cells, prostate epithelial cells and prostate cancer tissues, which may play multiple roles in prostate cancer progression and metastasis.

Overexpression of Notch3 and pS6 Is Associated with Poor Prognosis in Human Ovarian Epithelial Cancer.

Notch3 and pS6 play important roles in tumor angiogenesis. To assess the expression of Notch3 and pS6 in Chinese ovarian epithelial cancer patients, a ten-year follow-up study was performed in ovarian epithelial cancer tissues from 120 specimens of human ovarian epithelial cancer, 30 specimens from benign ovarian tumors, and 30 samples from healthy ovaries by immunohistochemistry. The results indicate that the expression of Notch3 and pS6 was higher in ovarian epithelial cancer than in normal ovary tissues and in benign ovarian tumor tissues (p < 0.01). In tumor tissues, Notch3 expression and pS6 expression were negatively associated with age (p > 0.05) but positively associated with clinical stage, pathological grading, histologic type, lymph node metastasis, and ascites (p < 0.05 or p < 0.01). A follow-up survey of 64 patients with ovarian epithelial cancer showed that patients with high Notch3 and pS6 expression had a shorter survival time (p < 0.01), in which the clinical stage (p < 0.05) and Notch3 expression (p < 0.01) played important roles. In conclusion, Notch3 and pS6 are significantly related to ovarian epithelial cancer development and prognosis, and their combination represents a potential biomarker and therapeutic target in ovarian tumor angiogenesis.

Enhanced expression of LINE-1-encoded ORF2 protein in early stages of colon and prostate transformation.

LINE-1 (L1) retrotransposons are a source of endogenous reverse transcriptase (RT) activity, which is expressed as part of the L1-encoded ORF2 protein (L1-ORF2p). L1 elements are highly expressed in many cancer types, while being silenced in most differentiated somatic tissues. We previously found that RT inhibition reduces cell proliferation and promotes differentiation in neoplastic cells, indicating that high endogenous RT activity promotes cancer growth. Here we investigate the expression of L1-ORF2p in several human types of cancer.We have developed a highly specific monoclonal antibody (mAb chA1-L1) to study ORF2p expression and localization in human cancer cells and tissues.We uncover new evidence for high levels of L1-ORF2p in transformed cell lines and staged epithelial cancer tissues (colon, prostate, lung and breast) while no or only basal ORF2p expression was detected in non-transformed cells. An in-depth analysis of colon and prostate tissues shows ORF2p expression in preneoplastic stages, namely transitional mucosa and prostate intraepithelial neoplasia (PIN), respectively.Our results show that L1-ORF2p is overexpressed in tumor and in preneoplastic colon and prostate tissues; this latter finding suggests that ORF2p could be considered as a potential early diagnostic biomarker.

Significance of circadian gene Period2 expression in epithelial ovarian cancer and the effect of gene overexpression on growth of ovarian cancer xenografts in nude mice

To study the significance of circadian gene Period2 expression in epithelial ovarian cancer tissues and the effect of gene overexpression on the growth of ovarian cancer xenografts in nude mice. Twenty-two cases of ovarian cancer paraffin specimens in the First Hospital of Shanxi Medical University (ovarian cancer group) were chosed during Jau. 2010 to Dec. 2013, including 8 cases of stage I, 8 cases of stage II, and 6 cases of stage III, while 6 cases of benign ovarian epithelial tumor paraffin specimens were selected as control (benign tumor group). Period2 gene were detected by real-time quantitative PCR and western blot methods in different stages of ovarian cancer tumor tissues. Established the ovarian cancer xenografts in nude mice with ovarian cancer cell line SKOV3, and they were divided into 3 groups (n = 8), including the recombinant plasmid group, empty plasmid group and control group. Using gene transfection technique to transfer Period2 gene into tumor tissues, tested the expression of Period2 mRNA in tumor tissues by real-time quantitative PCR after transfection into all nude mice, monitored the transplant tumor growth and calculating the tumor inhibition rate, detected the antiapoptotic gene BRE, apoptosis related tumor necrosis factor receptor (TNFR1) and tumor suppressor gene NIX in tumor tissues by real-time PCR and western blot in different groups. (1) The expression level of Period2 mRNA in tumor tissues among ovarian cancer group stage I, II and III were respectively 2.59 ± 0.50, 0.47 ± 0.08 and 0.42 ± 0.08, but benign tumor group was 6.59 ± 1.05. The expression level of Period2 protein in ovarian cancer group stage I, II and III were respectively 0.835 ± 0.087, 0.412 ± 0.035 and 0.199 ± 0.031, while benign tumor group was 0.874 ± 0.094. The expression level of Period2 mRNA and protein in benign tumor group was higher than those in ovarian cancer group stage I, II or III (P < 0.01). With ovarian cancer stage increased, the expression of Period2 mRNA and protein were decreased or absent (P < 0.05). (2) Two weeks after transfection, the expression level of Period2 mRNA in recombinant plasmid group tumor tissue was significantly higher than those in the empty plasmid group or the control group (6.11 ± 0.56 vs 0.50 ± 0.09 vs 0.44 ± 0.08, respectively; P < 0.01), the transplanted tumor volume of recombinant plasmid group was significantly less than those in empty plasmid group or the control group [(486 ± 70) mm(3) vs (835 ± 106) mm(3) vs (846 ± 110) mm(3), respectively; P < 0.01], the tumor inhibition rate of the recombination plasmid group was as high as 42.9%, that was significantly higher than those in the empty plasmid group and the control group (3.8% and 0, respectively; P < 0.05). (3) The expression level of BRE mRNA and protein in transplanted tumor tissues in the recombinant plasmid group were significantly lower than those in empty plasmid group and the control group; the expression level of TNFR1 and NIX were significantly higher than those in the empty plasmid group and the control group (all P < 0.05). Period2 mRNA and protein expression are absent in ovarian cancer of advanced stage. Transfection and stable expression of Period2 gene could slow down the growth of ovarian cancer, and the tumor inhibition rate could be significantly increased. Period2 gene may promote ovarian cancer cells apoptosis through inhibition of BRE gene expression and promoting TNFR1, NIX gene expression to exert anti-tumor effect.

Overexpression of eukaryotic translation initiation factor 5A2 (EIF5A2) correlates with cell aggressiveness and poor survival in gastric cancer.

Eukaryotic translation initiation factor 5A2 (EIF5A2) plays an important role in tumor progression and prognosis evaluation. However, little information is available about its potential role in gastric cancer. This study aimed to investigate the function of EIF5A2 in tumor progression and its potential mechanisms. EIF5A2 expression was measured in human gastric cancer cell lines, the immortalized gastric mucosal epithelial cell line (GES-1) and human gastric cancer tissues and knocked down by RNA interference or upregulated by EIF5A2 plasmid transfection. Cell proliferation, migration and invasion were assessed in vitro. The downstream targets of EIF5A2 were examined by western blotting. EIF5A2 and its potential target metastasis-associated protein 1 (MTA1) expression were examined in 160 pairs of human gastric cancer and adjacent non-tumor specimens using immunohistochemistry (IHC) staining, and its correlation with clinicopathological features and survival was investigated. Knockdown of EIF5A2 or MTA1 caused an apparent suppression of HGC27 cell proliferation, migration and invasion. After knockdown of EIF5A2 in HGC27 cells, E-cadherin levels were upregulated and vimentin, cyclin D1, cyclin D3, C-MYC and MTA1 levels were downregulated. Upregulation of EIF5A2 in MKN45 cells resulted in the converse. IHC results showed a positive correlation between EIF5A2 and MTA1 expression in gastric cancers (P<0.001). Both EIF5A2 and MTA1 overexpression were correlated with pT stage (P=0.018 and P=0.042), pN stage (P=0.037 and P=0.020) and lymphovascular invasion (P=0.016 and P=0.044). EIF5A2 or MTA1 overexpression was significantly associated with poor overall survival and disease-free survival (All P<0.05). Multivariate analyses identified EIF5A2 as an independent predictor for both overall survival (P=0.012) and disease-free survival (P=0.008) in gastric cancer patients. Our findings indicate that EIF5A2 upregulation plays an important oncogenic role in gastric cancer. EIF5A2 may represent a new predictor for poor survival and is a potential therapeutic target for gastric cancer.

B7-H1和Bcl-2在上皮性卵巢癌中表达及其临床意义

Objective To investigate the expressions of B7-H1 and Bcl-2 proteins in epithelial ovarian cancer, and to explore the association of the expressions of B7-H1 and Bcl-2 with clinicopathological features. Methods The expressions of B7-H1 and Bcl-2 proteins were detected by immunohistochemistry in 80 cases of epithelial ovarian cancer tissues and 20 normal ovary tissues. The association of the expressions of B7-H1 and Bcl-2 with the clinicopathological features was analyzed. Results The positive expression rates of B7-H1 and Bcl-2 in epithelial ovarian cancer tissues were 77.5% (62/80) and 58.8% (47/80), both higher than 15.0% (3/20) and 10.0% (2/20) in normal ovary tissues with significant difference (χ2=27.473, P<0.05; χ2=15.216, P<0.05). Both of Bcl-2 and B7-H1 expressions in epithelial ovarian cancer tissues were negatively correlated with the differentiation degree of epithelial ovarian cancer (χ2=9.367, P<0.01; χ2=11.702, P<0.01). The Bcl-2 expression in epithelial ovarian cancer tissues was positively correlated with the FIGO stage (χ2=7.766, P<0.01). The expression of B7-H1 was positively correlated with the expression of Bcl-2 in epithelial ovarian cancer (r=0.400, P<0.01). Conclusion The expressions of B7-H1 and Bcl-2 are up-regulated in epithelial ovarian cancer and they correlate to each other positively. The expressions of B7-H1 and Bcl-2 are correlated with the invasion and metastasis of epithelial ovarian cancer. The detection of B7-H1 combined with Bcl-2 may have an important clinical significance in the diagnosis and treatment for patients with epithelial ovarian cancer. Key words: Ovarian neoplasms; Gene; B7-H1; Bcl-2; Immunohistochemistry