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Objective:To investigate the effect of lncRNA AC132217.4 on the proliferation and invasion of liver cancer MHCC97-H cells and its molecular mechanism.Methods:The TCGA database was used to analyze the differential expression of AC132217.4 in liver cancer tissue and adjacent tissue, and to analyze the relationship between the expression level of AC132217.4 and the overall survival of liver cancer patients. Transfection of pcDNA-AC132217.4 plasmid into MHCC97-H cells was defined as AC132217.4 group, transfection of pcDNA plasmid into MHCC97-H cells was defined as negative control (NC) group, respectively. The proliferation and invasion ability of MHCC97-H cells were detected by MTT method and Matrigel invasion assay. The binding site between AC132217.4 and miR-18a-5p was analyzed by starBase v2.0 software and dual luciferase reporter gene assay. Real-time quantitative PCR (RT-qPCR) detected the differential expression of miR-18a-5p in the two groups of MHCC97-H cells. The expression of epithelial-mesenchymal transition protein was detected by Western-blotting. Measurement data with normal distribution were expressed as mean±standard deviation ( ± s), and t-test was used for comparison between the two groups. Results:Compared with adjacent tissues, the expression of AC132217.4 was down-regulated in liver cancer tissues ( P<0.01). Compared with liver cancer patients with low expression of AC132217.4, the overall survival of liver cancer patients with high AC132217.4 expression was longer ( P<0.05). The pcDNA-AC132217.4 plasmid significantly inhibited the proliferation of MHCC97-H cells ( P<0.05). The number of invasive cells in the NC group and AC132217.4 group were (131.30±12.55) and (37.45±7.77), respectively. The pcDNA-AC132217.4 plasmid significantly inhibited the invasive ability of MHCC97-H cells ( t=6.36, P<0.01). AC132217.4 directly complemented miR-18a-5p ( P<0.01). The expression of miR-18a-5p in MHCC97-H cells in AC132217.4 group (1.04±0.30) was significantly lower than that in NC group (6.13±0.75) ( t=6.27, P<0.01). Compared with the NC group, the expressions of epithelial phenotype proteins Cytokeratin and Claudin-1 in MHCC97-H cells in AC132217.4 group were up-regulated, while the expressions of mesenchymal phenotype proteins Vimentin, Slug and Snail were down-regulated. Conclusions:The expression of AC132217.4 is low in liver cancer tissue, and it is related to the overall survival of liver cancer patients. AC132217.4 might inhibit the proliferation and invasion of liver cancer MHCC97-H cells by sponge miR-18a-5p.
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Objective To explore the predictive value of the expression of CD44v6 and EGFR on the efficacy of neoadjuvant chemotherapy (NACT) in stageⅡ-Ⅲ cervical cancer. Methods A total of 53 patients with stageⅡ-Ⅲ cervical cancer diagnosed by pathology were selected. All patients received two cycles of paclitaxel+platinum NACT. The pathological tissue samples of cervical tumors before NACT treatment were collected. The expression of CD44v6 and EGFR were detected by the immunohistochemical SP method, and we analyzed their predictive value of NACT in stageⅡ-Ⅲ cervical cancer. Results Among the 53 patients, 38 were in the NACT effective group (CR+PR), and 15 were in the NACT ineffective group (SD+PD). The expression of CD44v6 in the ineffective group was significantly higher than that in the effective group (P < 0.05). The expression of CD44v6 was significantly different in patients with CR, PR, and SD (P < 0.05). The AUC of CD44v6 to NACT effect on stage Ⅱ-Ⅲ cervical cancer was 0.74 (P < 0.05). The patients in the high expression group of CD44v6 had worse efficacy in NACT than those in the low expression group of CD44v6 (P < 0.05). Pearson test showed that CD44v6 and EGFR expression were correlated (R=0.34, P < 0.05). Conclusion High expression of CD44v6 may reduce the efficacy of NACT in stageⅡ-Ⅲ cervical cancer, suggesting that the expression of CD44v6 has a certain predictive value and clinical significance in the efficacy of paclitaxel+platinum NACT on cervical cancer. Moreover, CD44v6 is positively correlated with EGFR expression.
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Objective To observe the effect of microRNA-613 (miR-613) on the biological behavior of human renal cell carcinoma cells and explore its molecular mechanism.Methods The synthesized miR-613 mimics and miR-NC were respectively transferred into 786-0 cells by Lipofectamine 3000 and divided into experimental group and control group.Real time quantitative polymerase chain reaction (qPCR) was used to detect the expression of miR-613 in cells of two groups to verify the transfection success.The proliferation of 786-0 cells was detected by methyl thiazolyl tetrazolium (MTT) assay and the migration of 786-0 cells was detected by Transwell migration assay.Bioinformatics predicts the target gene of miR-613.Dual luciferase reporter assay validates the targeted relationship between miR-613 and target genes,and their binding sites.qPCR and Western blot were used to detect the expression of target genes.Results qPCR test results showed that the expression of miR-613 in the experimental group (16.22 ± 1.08) was significantly higher than that in the control group (1.06 ± 0.20),with statistically significant difference (P ＜ 0.01).The results of MTT showed that the cells in the experimental group showed a decrease in absorbance value at the d-th point after transfection (P ＜ 0.05).Transwell cell migration test results showed that the cells in the control group and experimental group migrated from the upper chamber of Transwell to the lower chamber of the cells were respectively (95.55 ± 17.88) and (199.10 ± 22.74),with statistically significant difference (P ＜ 0.05).Bioinformatics predicts that the target gene of miR-613 is Cortactin (CTTN).Dual luciferase reporter assay confirmed that miR-613 can target CTTN gene (P ＜ 0.05).miR-613 can significantly inhibit the CTTN gene expression in renal cell carcinoma cells (P ＜ 0.01).Conclusions Upregulation of miR-613 expression can inhibit the proliferation and migration of renal cell carcinoma cells,and its possible molecular mechanism is that miR-613 inhibits the expression of CTTN gene.
