Silencing Research Articles

The correlation of ESCO1 expression with a prognosis of prostate cancer and anti-tumor effect of ESCO1 silencing

Background: Recently, it has been reported that establishment of sister chromatid cohesion N-acetyltransferase 1 (ESCO1) is involved in tumorigenesis. However, its role in prostate cancer remains unclear. In the present study, the association between ESCO1 expression and the prognosis of prostate cancer was investigated, and the potential molecular mechanisms underlying its actions in tumor progression were also examined. Methods: Immunohistochemical analysis was performed to detect the expression of ESCO1 in benign prostatic hyperplasia (BPH), human prostate cancer, and metastasis tissue samples, and the association between the establishment of ESCO1 expression and the prognosis of prostate cancer was investigated. The effect of ESCO1 expression on the viability, migration, and invasion of prostate cancer cells in vitro was analyzed, along with the effect of ESCO1 silencing on the growth of prostate tumors in vivo . Results: The results demonstrated an increase in the expression of ESCO1 in prostate cancer tissue when compared with BPH, and it was significantly associated with tumor malignancy and poor patient survival. Additionally, knockdown of ESCO1 significantly inhibited the viability and migration of prostate cancer cell. Furthermore, we found that knockdown of ESCO1 significantly inhibited tumor growth in vivo . Pathway analysis identified that the silencing of ESCO1 significantly decreased the phosphorylation levels of protein kinase B. Conclusions: The results of the present study indicate that ESCO1 plays a vital role in the progression of human prostate cancer; furthermore, ESCO1 may potentially serve as a prognostic marker and a novel therapeutic target for this disease.

BMP7 Gene Silencing Inhibits Osteogenic Differentiation of Porcine Arotic Valve Interstitial Cells Induced by Osteogenic Induction Medium

BMP7 Gene Silencing Inhibits Osteogenic Differentiation of Porcine Arotic Valve Interstitial Cells Induced by Osteogenic Induction Medium

Self-assembling Nanotechnology for Cancer Personalized Medicine

domenico.marson@dia.units.itTheranostics is a new field of medicine, which combines specific targeted therapies and diagnostic tests. With a key focus on patient centered care, theranostics provides a transition from conventional medicine to a contemporary personalized and precision medicine approach. The theranostic paradigm in cancer involves nanoscience to unite diagnostic and therapeutic applications to form nanosized agents for diagnosis, drug/gene delivery and treatment response monitoring. These nanocarriers can indeed be engineered to precisely control drug/gene/sensor-release rate and/or target specific organs/tissues within the body with a specific amount of therapeutic/diagnostic agent. In order to fulfill these expectations, any nanovector system must be designed to transport the optimum amount of therapeutic/diagnostic cargo to the desired target site where the active principle is to be released at an optimal rate during a specific time window.Keeping the promises of theranostics is a current formidable challenge in (bio)nanotechnology, and major efforts are devoted to the design of integrated multifunctional and multivalent nanovectors able to provide selective recognition combined with sustained release and/or diagnostic reporting. In this contribution, the pathway leading to two types of nanosystems obtained by exploiting the quintessence of nanotechnology, i.e., the self-assembling process of small, amphiphilic molecules, is reported. Depending on the specific chemistry adopted, these nanomicelles are able to perform specific and effective gene silencing via targeted small interfering RNA (siRNA) delivery, and provide PET images with significantly superior imaging quality relating to sensitivity, specificity and accuracy when compared to the clinical standard [18F]FDG.

Silencing ANO9 increases the radiosensitivity of pancreatic cancer cell AsPC-1

Objective To evaluate the effect of ANO9 on the radiosensitivity of pancreatic cancer cell AsPC-1, aiming to provide new targets for clinical radiotherapy of pancreatic cancer. Methods Western blot was performed to detect the expression of ANO9 in pancreatic cancer cell lines (BxPC-3, PANC-1, AsPC-1) and normal pancreatic cell line (HPNE). The AsPC-1 cell line with stable silencing ANO9 was constructed by using lentivirus and validated by Western blot. MTT assay was adopted to detect the cell viability of AsPC-1 with stable silencing ANO9 after irradiation. Colony formation assay was conducted to evaluate the effect of silencing ANO9 upon the radiosensitivity of AsPC-1 cells. Western blot was performed to assess the effect of ANO9 silencing on the expression of EGFR/ERK signaling protein. Results The expression levels of ANO9 were significantly up-regulated in three pancreatic cancer cell lines compared with that in the normal pancreatic cell line HPNE (t=7.426, 5.543, 11.850, all P<0.05). After silencing ANO9, the expression level of ANO9 protein was significantly down-regulated than that in the control group (t= 9.670, P<0. 05). The AsPC-1 cells with stable silencing ANO9 were successfully constructed. The sensitivity of AsPC-1 cells to irradiation was significantly increased after silencing ANO9, and the sensitivity enhancement ratio was 1.566. The expression levels of EGFR/ERK signaling proteins (EGFR and p-ERK1/2) were significantly down-regulated after silencing ANO9(t=7.949, 13.160, both P<0.05). Conclusions Silencing ANO9 can significantly increase the sensitivity of AsPC-1 cells to radiotherapy, which is probably associated with the inhibition of EGFR/ERK signaling transduction. ANO9 might be a new therapeutic target for preventing the progression of pancreatic cancer. Key words: ANO9 gene; Pancreatic cancer cell line; Radiosensitivity

AmiRNA-mediated PL Gene Silencing Driven by the Fruit-specific Promoter P119 in Strawberry and Its Effect on Fruit Firmness

amiRNA-mediated PL Gene Silencing Driven by the Fruit-specific Promoter P119 in Strawberry and Its Effect on Fruit Firmness

Corrigendum: miR156/SPL10 Modulates Lateral Root Development, Branching and Leaf Morphology in Arabidopsis by Silencing AGAMOUS-LIKE 79

[This corrects the article DOI: 10.3389/fpls.2017.02226.].

Effect of low expression of tripartite motif-containing protein 28 on Hep3B proliferation

Objective To investigate the effect of low expression of tripartite motif-containing protein 28 (TRIM28) and downstream target protein β-catenin. Methods TRIM28 was silenced in Hep3B cells mediated by lentiviral vector, and the proliferation of Hep3B cells after TRIM28-knockdown was detected by cell colony. Results After TRIM28 knockdown, the protein level of β-catenin was significantly down-regulated, which was 0.78 of the control group, and β-catenin/glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in the KD1 group and KD2 group decreased to 0.28 and 0.25, respectively (P<0.05); The proliferation of Hep3B hepatoma cells was significantly inhibited (P<0.05). Conclusion TRIM28 has a promoting effect on the proliferation of Hep3B hepatoma cells. Key words: Hepatocarcinoma; Tripartite motif-containing protein 28; β-catenin

Effect of silencing troponin I3 gene expression on biological property of rat embryonic H9C2 cardiomyocytes

Objective To investigate the effect of silencing troponin I3 (Tnni3) gene expression on biological property of rat embryonic H9C2 cardiomyocytes. Methods The rat embryonic H9C2 cardiomyocytes were cultured and divided into 2 groups: control group transfected with negative control small interfering RNA (NC-siRNA group) and experimental group transfected with Tnni3 small interfering RNA (Tnni3-siRNA group). At 48 h, 72 h after transfection, the cells were collected, and real time quantitative polymerase chain reaction (qPCR)was used to detect the mRNA expressions of Tnni3 and Caspase-3, and Western blot was used to detect the protein expressions of Tnni3, Cyclin A1 and Cyclin B1.Annexin V-fluorescein isothiocyanate(FITC) apoptosis detection kit was used to analyze cell apoptosis.Cell proliferation was measured by Cell Counting Kit-8 (CCK-8) solution and cell cycle was detected by flow cytometry. Results At 48 h post-transfection with Tnni3-siRNA, H9C2 cells exhibited a significant decrease in Tnni3 mRNA (0.27±0.05 vs. 1.00±0.00) and protein (0.18±0.03 vs. 1.00±0.00) compared with those transfected with NC-siRNA, and the differences were statistically significant (t=25.26, 47.40, all P<0.01). Apoptotic cells were observed in the NC-siRNA group and the Tnni3-siRNA group.At 72 h post-transfection, the percentage of apoptotic cells significantly increased in H9C2 cells transfected with Tnni3-siRNA [(11.30±1.85)% vs. (0.33±0.15)%] compared with those transfected with NC-siRNA, an increased expression of Caspase-3 mRNA was also observed in Tnni3-siRNA-transfected H9C2 cells (1.39±0.13 vs. 1.00±0.00), and the differences were statistically significant (t=10.24, 5.19, all P<0.01). Compared with NC-siRNA-transfected H9C2 cells, a time-dependent reduction in cell proliferation was observed in Tnni3-siRNA-transfected H9C2 cells (48 h: 0.32±0.06 vs. 0.46±0.03; 72 h: 0.31±0.01 vs. 0.63±0.04; 96 h: 0.36±0.01 vs 0.75±0.04), and the differences were statistically significant (t=3.62, 13.45, 16.39, all P<0.01). At 72 h post-transfection with Tnni3-siRNA, the percentage of G1 phase, S phase and G2 phase cells was (71.25±3.82)%, (18.28±2.78)% and (9.94±1.09)%, respectively.There was a significant increase in the proportion of G2 phase cells [(9.94±1.09)% vs. (4.54±0.99)%] in H9C2 cells transfected with Tnni3-siRNA compared with those transfected with NC-siRNA, an increased expression of Cyclin A1 protein (1.89±0.09 vs.1.00±0.00) and a decreased expression of Cyclin B1 protein (0.47±0.06 vs.1.00±0.00) were observed in Tnni3-siRNA-transfected H9C2 cells, respectively, and the differences were statistically significant (t=6.35, 17.12, 15.32, all P<0.01). Conclusions Silencing Tnni3 gene expression in rat embryonic H9C2 cardiomyocytes can induce cell apoptosis, suppress cell proliferation, and led to G2 cell cycle arrest. Key words: Troponin I3 gene; Cell apoptosis; Cell proliferation; Cell cycle

Role of LncRNA TCONS_00003757 in papillary thyroid carcinoma

Objective To investigate the expression of LncRNA TCONS_00003757 in papillary thyroid carcinoma(PTC), and to explore the effect of LncRNA TCONS_00003757 on the proliferation, migration, and invasion of TPC-1 cells. Methods Sixty cases of papillary thyroid carcinoma specimens and paired normal tissues were collected after surgery from October 2017 to March 2018 in the Tangshan Workers Hospital. Real-time PCR analysis was used to examine the expression level of LncRNA TCONS_00003757 in PTC tissues, and the association of LncRNA TCONS_00003757 expression levels with the clinicopathological data of patients was analyzed. The effects of LncRNA TCONS_00003757 and ST3GAL5 silencing on the proliferation, migration, and invasion of TPC-1 cells were assessed by MTT, scratch test, and transwell assay. Protein expression of ST3GAL5 was detected by Western blot after LncRNA TCONS_00003757 silencing. Results LncRNA TCONS_00003757 expression was increased in PTC tissues compared with that in normal tissues(P<0.05), which was correlated with capsule invasion and lymph node metastasis(P<0.05). LncRNA TCONS_00003757 expression in TPC-1 cells was higher than that in normal thyroid follicular epithelial Nthy-ori 3-1 cells(P<0.05). LncRNA TCONS_00003757 and ST3GAL5 silencing inhibited the proliferation, migration, and invasion of thyroid cancer TPC-1 cells, and ST3GAL5 expression was decreased after LncRNA TCONS_00003757 silencing(P<0.05). Conclusion LncRNA TCONS_00003757 expression in PTC is significantly increased and correlated with its occurrence and development. Silencing LncRNA TCONS_00003757 may inhibit the proliferation, migration, and invasion of thyroid papillary cancer cells via decreasing ST3GAL5 expression. Key words: Thyroid papillary carcinoma; LncRNA TCONS_00003757; Long non-coding RNA

LncRNA CCAT1 enhances radiosensitivity of human pancreatic cancer cells PANC-1 by targeting miR-130b-3p

Objective To investigate the effect of lncRNA CCAT1 and miR-130b-3p on the radiosensitivity of human pancreatic cancer cells PANC-1. Methods Real-time PCR was used to detect the relative expression levels of CCAT1 and miR-130b-3p in pancreatic cancer tissues and cell lines including PANC-1 cells irradiated with 2 Gy X-rays. After silencing CCAT1 and/or inhibiting miR-130b-3p expression, cell apoptosis rate, Caspase 3 activity and cell survival were detected by flow cytometry, Caspase 3 activity detection kit and colony formation assay, respectively. Cell survival curve was stimulated by the multi-target single-hit model. Based on the starBase v2.0 online analysis, the luciferase reporter gene assay, RNA-binding protein immunoprecipitation assay (RIP) and Real-time PCR assay were applied to verify the relationship between CCAT1 and miR-130b-3p. Results CCAT1 expression was up-regulated (t=6.322-8.555, P<0.05), but miR-130b-3p expression was down-regulated (t=3.950-18.795, P<0.05) in the radiation-resistant pancreatic cancer tissues, pancreatic cancer cell lines and 2 Gy-irradiated PANC-1 cells. When the CCAT1 silenced PANC-1 cells were irradiated with 2 Gy, cell survival fraction decreased (t=2.929, 5.047, 5.234, 5.125, P<0.05), apoptosis rate and Caspase 3 activity increased (t=6.953, 6.836, P<0.05). CCAT1 could selectively regulate miR-130b-3p expression. Inhibition of miR-130b-3p expression could enhance PANC-1 cell survival (t=4.564, 6.736, 8.656, P<0.05), but reduced apoptosis rate (t=5.234, P<0.05) and Caspase 3 activity (t=10.440, P<0.05). Conclusions Silencing CCAT1 promotes the expression of miR-130b-3p and enhances radiosensitivity of PANC-1 cells. Key words: lncRNA CCAT1; miR-130b-3p; Pancreatic cancer; Radiosensitivity

Circular RNA expression profiles in women with gestational diabetes mellitus: a nested case-control study

Objective To investigate the expression of circular RNA (circRNA) in peripheral blood of gravidas with gestational diabetes mellitus (GDM) using gene chip technology, and to provide evidence for studying pathogenesis of GDM. Methods A prospective cohort-based nested case-control study was used to select 1 018 pregnant women who were examined and delivered in Guangzhou Women and Children's Medical Center from September 2014 to April 2016. The information of prenatal examination was recorded and the outcome of delivery was followed up. Six pregnant women diagnosed as GDM were selected as the GDM group. Six normal pregnant women who were collected blood samples by 1∶1 were selected as the control group. The difference of gestational week was less than 7 days, the number of previous pregnancies was less than 2 times, and the difference of previous delivery times as same as the control group. The expression of circRNA in maternal peripheral blood was analyzed by chromosome microarray technique, and the function and the regulation network forecast were analyzed by GO (http://www.geneontology.org/), KEGG PATHWAY (http://www.genome.jp/kegg/pathway.html) and CircNet(http://circnet.mbc.nctu.edu.tw/). t-test was used for data analysis. Results Compared to the normal pregnant women, 2 678 circRNAs were identified to be differentially expressed >2.0 times in GDM women, among which 1 532 were up-regulated and 1 146 were down-regulated. Functional analysis showed that the up-regulated circRNA was enriched in the biological processes of insulin response, gene silencing regulation, glucagon response and cell senescence. Signal pathway analysis showed that circRNA involved in insulin pathway. Taking has_circ_0042852 and has_circ_0004001 as center, the possible GDM-related regulatory networks were predicted. Conclusions The peripheral blood of GDM women is rich in circRNAs, which might involve in many biological processes, insulin signaling pathway and possibly induding GDM-related regulatory networks. Key words: Diabetes, gestational; RNA; Gene expression profiling; Biomarkers

Improved agonists for chemogenetics

Neuroscience Targeting ligand-responsive receptors to specific groups of cells, a strategy known as chemogenetics, is a powerful tool in many neurological applications. There is increasing interest in extending these tools for human treatment. Magnus et al. designed chemogenetic ion channels that improve currently available systems and are activated by the clinically used antismoking drug varenicline. They engineered a ligand-binding domain less responsive to endogenous signals and identified agonists that function at nanomolar concentrations. The combination of drug and introduced channels transiently silenced neurons, with slow but effective washout, and induced behavioral changes in animal models after brain administration. Science , this issue p. [eaav5282][1] [1]: /lookup/doi/10.1126/science.aav5282

ShRNA-mediated silencing of hTERT promote apoptosis and senescence in human ovarian cancer cells

Background: Human telomerase reverse transcriptase (hTERT) is a crucial oncogene and a key factor in cell immortalization. Apart from hematopoiesis stem cell and germ cells, normal cells and tissues almost no expression of this gene. However, the positive expression rate of hTERT in malignant tumors is as high as 80–95%. This study was designed to investigate the mechanism of knockdown of hTERT expression in the regulation of ovarian cancer carcinogenesis in nude mice transplantation model. Methods: We identified the function of hTERT in the treatment of ovarian cancer by constructing a nude mice transplantation tumor model. Then analyzed the mechanism of hTERT in modulating ovarian cancer cell apoptosis and senescence by immunohistochemically staining. Results: Our analysis identified hTERT as a novel regulator of ovarian cancer progression and is one of the most significantly upregulated genes in ovarian cancer. Down-regulation of hTERT inhibits proliferation of ovarian cancer cells by promoting apoptosis and the expression of the senescence gene p53 and p21 . Conclusions: hTERT is a key oncogene in ovarian tumorigenesis and metastasis; the downregulation of hTERT significantly inhibit the proliferation of ovarian cancer and have an effect on treatment of ovarian cancer by activating cell apoptosis and senescence pathway.

Effect of microRNA-19b on proliferation, migration and invasion of pancreatic ductal adenocarcinoma

Objective To examine the expression of microRNA (miRNA, miR)-19b in pancreatic ductal adenocarcinoma (PDAC) cell lines, and the functional role of miR-19b in proliferation, migration and invasion of PDAC cells. Methods Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to detect the expression level of miR-19b in human pancreatic ductal epithelial cell line (HPDE), human PDAC cell lines (PANC1, MiaPaCa2 and BxPC3) and huamn PDAC metastasis cell line (COLO357) and its fast-growing variant FG. The expression of miR-19b was inhibited or overexpressed in PDAC cell lines. Transwell assay and Matrigel assay were carried out to examine the migration and invasion capabities of PDAC cells with miR-19b silencing or overexpressing. Methyl thiazol tetrazolium (MTT) and colony formation assays were performed on PDAC cell lines with miR-19b silencing or overexpression. Western blotting was used to detect the expression of proliferation markers (p21 and p27) in PDAC cell lines after miR-19b was silenced or overexpressed. Results The results of RT-qPCR showed the expression level of miR-19b in human PDAC cell lines PANC1, MiaPaCa2, BxPC3 and metastatic cell line COLO357 and FG (198.45±21.22, 221.37±18.59, 182.51±13.48, 317.64±15.20, 331.34±17.18) was significantly higher than that in normal human pancreatic ductal HPDE (98.21±14.54, P<0.05). Transwell cell migration assay and Matrigel cell invasion assay showed that the migration and invasion ability of FG cells after miR-19b silencing [(22.54±2.87)%, (28.87±5.62)%] was significantly decreased as compared with the control group [(99.65±3.07)%, (98.73±4.38)%, P<0.01]. In contrast, the migration and invasion ability of BxPC3 cells overexpressing miR-19b [(317.53±10.21)%, (286.77±12.83)%] was significantly higher than that of the control group [(97.89±5.31)%, (98.25±7.57)%, P<0.01]. The colony formation and MTT assays showed that the proliferation ability of PDAC cells after miR-19b was silenced was significantly decreased as compared with the control group (P<0.01). The result of Western blotting showed that the expression of proliferation markers (p21 and p27) in PDAC was increased by (3.25±0.27) and (5.38±0.33) times respectively as compared with the control group after miR-19b silencing (P<0.01). When miR-19b was overexpressed, the expression of p21 and p27 decreased by (2.68±0.15) and (3.27±0.27) times respectively as compared with the control group (P<0.01). Conclusion MiR-19b could promote the migration and invasion of PDAC cells and increase the proliferation capability. Key words: Pancreatic carcinoma; MicroRNA-19b; Metastsis; Proliferation

Effect of ZNF593 gene on proliferation of glioma cells

Objective To investigate the expression level of ZNF593 in glioma cells and its effect on the proliferation of glioma cells. Methods The date from TCGA were analyzed to explore the expression of ZNF593 gene in glioma cells and its relation to prognosis. Glioma U251 cells were transfected with ZNF593 short hairpin RNA (shZNF593) and shRNA control (shCtrl) respectively. The mRNA expression of ZNF593 in the cells was detected by real-time quantitative polymerase chain reaction (Real-time PCR), and the protein level of ZNF593 was measured by Western blotting. Cell proliferation was detected by Celigo and methyl thiazol tetrazolium (MTT). Results The date from TCGA shows ZNF593 gene was highly expressed in glioma, and high expression of ZNF593 leads lower survival rate. After the ZNF593 gene was silenced, Real-time PCR showed that the expression abundance level of shZNF593 was 0.380, and that was 1.000 of shCtrl, the knockdown efficiency was 62%; Western blotting showed that the protein level of ZNF593 was decreased; Celigo Cell Counting indicated that the daily increase from the second day of shCtrl group were 1.971, 2.788, 3.827, 5.169, and those of shZNF593 group were 1.340, 1.641, 2.026, 2.095; The result of MTT in 490 nm from second day to fifth day of shCtrl group were 0.174, 0.316, 0.485, 0.620, 0.765, and those of shZNF593 group were 0.172, 0.235, 0.268, 0.316, 0.366 (P<0.05). Conclusion ZNF593 is significantly up-regulated in glioma, knock down ZNFF593 can suppress the proliferation of glioma cells. Key words: ZNF593; Glioma; Cell proliferation

Research progress of peripheral blood regulatory T cell/helper T cell 17 imbalance and relevant gene methylation in immune thrombocytopenia

Immune thrombocytopenia (ITP) is an autoimmune hemorrhagic disease. The classic pathogenesis of ITP is that the production of autoantibodies against platelet membrane glycoproteins, which mediates platelet destruction. In recent years, it has been found that there are still many other mechanisms in ITP inducing thrombocytopenia and platelet damage, among which abnormal cellular immunity plays an important role. In cellular immunity, CD4+ CD25high CD127- regulatory T cells (Treg) are a group of T cell subsets that exist in human peripheral blood and spleen and can inhibit auto-reactive T cell responses; T helper cells (Th)17 is a newly discovered T cell subset that secretes interleukin (IL)-17, involved in mediating inflammatory response, which is of great significance in autoimmune diseases. Several studies showed that Treg/Th17 imbalance and abnormal expression of specific transcription factors folkhead box protein (Foxp)3 and retinoic acid-related orphan nuclear receptor (ROR)-γt were observed in peripheral blood of ITP patients. DNA methylation induces Foxp3 gene silencing and abnormal activation of IL-6/STAT3 signaling pathway, which may be the mechanism leading to Treg/Th17 imbalance and causes of ITP. The proposed demethylation treatment may provide new ideas for the molecular targeted treatment of ITP, but its scientificity, effectiveness and safety still need to be confirmed by further studies. The author reviews literature on the role of Treg/Th17 imbalance and relevant gene methylation mechanism in the occurrence, development and treatment of ITP. Key words: T-lymphocytes, regulatory; T-lymphocyte subsets; DNA methylation; Thrombocytopenia; Helper T lymphocyte 17; Decitabine

Impact of NANOG gene on the invasion ability of bladder cancer cell line

Objective To evaluate the influence on the invasion ability of T24 bladder cancer cell lines after silencing of NANOG gene. Methods Human urinary bladder cancer cells T24 were cultured and transfected with short hairpin RNA (shRNA) to silence NANOG gene. Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) and enzyme linked immunosorbent assay (ELISA) were performed to detect the expression level of matrix metalloproteinase (MMP)-2, MMP-9 and inhibitor of matrix metalloproteinases-1 (TIMP-1). Wound-healing assay and invasion test were performed to determine the invasion capability of T24 cell lines. Results When NANOG gene was silenced, the expression of MMP-2 and MMP-9 was down-regulated (for MMP-2 by 63.21%, P<0.05; for MMP-9 by 73.55%, P<0.05). ELISA showed that the expression of MMP-9 in supernatant of T24 cells was down-regulated by 21.07% (P<0.05). Scratch test and invasive test showed that the invasive ability of cells decreased after NANOG gene was silenced (all P<0.05). Conclusion NANOG may modulate the invasion of bladder cancer cells through regulating the expression of MMP-2 and MMP-9. Key words: Bladder cancer; Metal matrix proteinase 2; Metal matrix proteinase 9; Short hairpin RNA; Invasion

In vitro and in vivo effects of RNA interference targeting c-Myc gene on gastric cancer SGC7901 cells

Objective To observe the effect of RNA interference to c-Myc on proliferation of gastric cancer SGC7901 cells in vitro and in vivo. Methods The c-Myc gene of gastric cancer SGC7901 was silenced by RNA interference technique. The expression of c-Myc gene of SGC7901 was detected by real-time fluorescent quantitative polymerase chain reaction (FQ-PCR). The proliferation of gastric cancer SGC7901 was detected by methyl thiazol tetrazolium (MTT) assay and in vivo tumorigenesis. Results After the silencing of c-Myc gene by RNA interference technology, the proliferation rate and tumorigenic growth rate of gastric cancer SGC7901 decreased significantly (on day 7, compared with the blank group, the tumor inhibition rate of small interfering RNA (siRNA)-scramble group was 98.78% (P>0.05), while that of the experimental group was 49.96% (P<0.05). Conclusion After the c-Myc interference, gastric cancer SGC7901 in vitro and in vivo proliferation rate decreased. Key words: Gastric carcinoma; RNA interference; c-Myc; Proliferation

Expression of special AT rich sequence binding protein-1 in lung adenocarcinoma and its effect on proliferation, invasion and apoptosis of lung adenocarcinoma cells

Objective The study aimed to explore the relationship between the expression of special AT rich sequence binding protein-1 (SATB1) and clinical characteristics in lung adenocarcinoma and its role in the proliferation, invasion and apoptosis of lung adenocarcinoma cell line A549. Methods Tissue sections were stained by immunohistochemistry streptavid-in-peroxidase (SP) method. The green fluorescent protein expression vector of SATB1-small interfering RNA (siRNA) was constructed and transfected into lung adenocarcinoma cell line A549. The effect of transfection was observed by fluor escence microscope. The silencing efficiency was measured by Western blotting. Cell proliferation, invasion and migration activity were detected by methyl thiazol tetrazolium (MTT), Transwell and wound-healing assay. Apoptosis rate was measured by flow cytometry. MTT, transwell and wound-healing assay were adopted to study cell proliferation, invasion and migration activity, and the apoptosis rate was tested by flow cytometry. Results SATB1 was highly expressed in human lung adenocarcinoma tissues (positive rate 61%), and was lowly expressed in adjacent tissues (positive rate 20%), and the expression level was negatively correlated with the degree of differentiation (P<0.01). After transfection with SATB1-siRNA, the transfection efficiency was about 70%, and the expression of SATB1 in A549 cells was decreased (P<0.01). Compared with the negative control group, the cell proliferation, invasion and migration ability were significantly decreased (P<0.01). The apoptotic rate was increased (P<0.01). Conclusion SATB1 is closely related to the pathogenesis and development of lung adenocarcinoma, which may provide important clues for finding potential therapeutic targets and prognosis of lung adenocarcinoma. Key words: Lung adenocarcinoma; Special AT rich sequence binding protein-1; Cell proliferation

Influence of proteasome activator γ on iodine uptake capability of anaplastic thyroid carcinoma

Objective To investigate the changes of iodine uptake capability (IUC) and mRNA expression of iodine uptake-related proteins in ARO and WRO thyroid carcinoma cell lines after silence of proteasome activator γ (REGγ), and observe the relation between REGγ and IUC of thyroid carcinoma. Methods The AROshN, AROshR, WROshN and WROshR (shN= blank plasmid, shR=plasmid with silence of REGγ) thyroid carcinoma cell lines were routinely cultured. Low dosage (3.7 kBq) of Na125I was added and then IUC was determined at different time points (5, 10, 15, 20, 40 and 70 min). The mRNA expressions of sodium/iodine symporter (NIS), thyroid stimulating hormone receptor (TSHR), thyroid peroxidase (TPO) and thyroglobulin (Tg) were examined by real-time PCR. Paired t test was used to analyze the data. Results After the silence of REGγ, the peak values of IUC in AROshR and WROshR cells were increased from (1 974±12) to (4 502±23) counts/min, and from (2 988±25) to (5 001±16) counts/min, respectively. The increase rates were 128.1% in AROshR cells and 67.4% in WROshR cells (t values: 17.30, 13.20, both P<0.05). The mRNA expressions of NIS, TSHR, TPO, Tg in AROshR cells were 2.82, 1.98, 2.65 and 2.31 times higher than those in AROshN cells, and the expressions in WROshR cells were 2.21, 1.78, 2.51 and 1.78 times higher than those in WROshN cells (t values: 13.80-21.93, all P<0.05). Conclusion Silence of REGγ can increase the gene expressions of the iodine uptake-related proteins and elevate the IUC of thyroid carcinoma cells. Key words: Thyroid neoplasms; Tumor cells, cultured; Proteasome endopeptidase complex; Gene silencing; Iodine radioisotopes