Long Non-coding RNA FAM230B-206 Regulates Cisplatin Resistance In Gastric Cancer Cell By MiR-532-3p/PAX6

Objective Gastric cancer is an important disease of death by malignant tumor,especially advanced gastric cancer,one of its important treatment is platinum-containing chemotherapy.In order to further improve the efficacy of cisplatin,it is particularly important to explore the emergence of drug resistance from the mechanism.Therefore,the Long non-coding RNA(Inc RNA)has become a new hot spot in tumor research.This topic explores the expression of IncRNA in gastric cancer cells,and focuses on the influence of IncRNA FAM230B-206 on gastric cancer cisplatin resistance and its regulatory mechanism.Methods The difference of IncRNA expression in gastric cancer cell line SGC7901/DDP and parental cell line SGC7901 was detected by microarray.FAM230B-206 with significant difference in screening was selected as the research object,the difference of FAM230B-206 expression was verified by RT-PCR,using lentivirus SGC7901/DDP cells stably over expression of FAM230B-206、stable knock down FAM230B-206 in SGC7901 cells.The gene expression changes after transfection were confirmed by RT-PCR,and the survival and IC50 of cells after cisplatin treatment were examined by CCK-8 assay to determine the change of drug resistance.Bioinformatics analysis FAM230B-206 and miR-532-3p have partial complementary pairing,and PAX6 is the common target of FAM230B-206 and miR-532-3p.The use of lentivirus over expression or knock down miR-532-3p,CCK-8 method to check changes in cell resistance.The expression of miR-532-3p after up-regulated or down-regulated by FAM230B-206 was detected by RT-PCR.Flow cytometry was used to analyze the effect of over expression or knock down FAM230B-206 on gastric cancer cell apoptosis after a certain period of time.Western blot was used to examine the expression of PAX6 after over expression or knock down FAM230B-206 and miR-532-3p,and to further infer the mechanism of FAM230B-206 affecting cisplatin resistance.Results The results of microarray showed that there were significant differences in the expression of 1077 Inc RNA in SGC7901/DDP cells and SGC7901 cells,and the expression of FAM230B-206 was significantly higher in SGC7901/DDP than in SGC7901(Fc=4.88),and the difference was statistically significant(p<0.01)may be a key molecule involved in cisplatin resistance in gastric cancer.Using lentiviral transfection and puromycin-selective RT-PCR validation,FAM230B-206 overexpression stably transfected cell line(OE-FAM230B-206)and FAM230B-206 low expressing stable transfectant cell line(KD-FAM230B-206)were established.And the corresponding airborne stable cell lines(NC-SGC7901 and NC-SGC7901/DDP).The CCK-8 cytotoxicity test showed that the IC50 of OE-FAM230B-206 was significantly higher than that of NC-SGC7901 and SGC7901,and the cell resistance to cisplatin was enhanced,while KD-FAM230B-206 was relative to NC-SGC7901/DDP and SGC7901/DDP.The IC50 decreased significantly,and the resistance of cells to cisplatin weakened.The difference was statistically significant.The apoptosis rate of cells treated with cisplatin for 48 hours by flow cytometry indicated that downregulation of FAM230B-206 expression resulted in a significant increase in the proportion of apoptosis of NC-SGC7901/DDP and SGC7901/DDP,upregulation of FAM230B-206 expression led to NC-SGC7901 and SGC7901 The proportion of apoptosis is significantly reduced.Bioinformatics analysis showed that FAM230B-206 and miR-532-3p have complementary pairings,and PAX6 is a common target for FAM230B-206 and miR-532-3p.In contrast to miR-532-3p expression in SGC7901/DDP and SGC7901 and FAM230B-206,miR-532-3p was down-regulated in SGC7901/DDP to reduce cisplatin resistance,and upregulation of FAM230B-206 expression in SGC7901 was resistant to cisplatin resistance.Medicinal improvement.Down-regulating the expression of FAM230B-206 in SGC7901/DDP cells increased the expression of miR-532-3p and decreased the expression of PAX6.In contrast,the expression of FAM230B-206 and miR-532-3p decreased and the expression of PAX6 increased in SGC7901 cells.Conclusion:lncRNA FAM230B-206 was significantly over-expressed in gastric cancer cisplatin-resistant cells.By knock down FAM230B-206 can significantly inhibit cisplatin resistance in gastric cancer,and over xpression of FAM230B-206 gastric cancer cells increased resistance to cisplatin.Its mechanism may be the regulation of gastric cancer cisplatin sensitivity involved in FAM230B-206 through miRNA-532-3p/PAX6.This study may provide a new idea for reversing the drug resistance of gastric cancer cisplatin and enhancing the sensitivity of chemotherapy.