Impacts of LINC00461 on proliferation,apoptosis,and cisplatin resistance of gastric cancer cells by regulating the miR-1301-3p/MYH9 axis
Objective To investigate the impacts of long non coding RNA LINC00461(LINC00461)on the proliferation,apoptosis,and cisplatin(CDDP)resistance of gastric cancer cells by regulating the miR-1301-3p/myosin heavy chain 9(miR-1301-3p/MYH9)axis,seeking new targets for the treatment of gastric cancer.Methods The reverse transcription-quantitative polymerase chain reaction(RT-qPCR)method was applied to detect the expression of LINC00461,miR-1301-3p,and MYH9 in normal gastric mucosal cells GES-1,gastric cancer cells HGC-27,and gastric cancer cisplatin resistant cell lines HGC-27/CDDP.Gastric cancer cisplatin resistant cells HGC-27/CDDP were separated into Control group,si-NC group,si-LINC00461 group,si-LINC00461+anti-NC group,and si-LINC00461+anti-miR-1301-3p group.The RT-qPCR method was applied to detect the expression of LINC00461,miR-1301-3p,and MYH9 in HGC-27/CDDP cells in each group.Edu experiment was applied to detect cell proliferation.Annexin V-FITHCA/PI was applied to detect cell apoptosis.Western blot was applied to detect the expression of cell proliferation and apoptosis related proteins.Dual luciferase assay was applied to detect the targeting relationship between LINC00461 and MYH9 with miR-1301-3p,respectively.Results In HGC-27/CDDP cells,the expression of LINC00461 and MYH9 mRNA was high,and the expression of miR-1301-3p was low;the differences were significant when compared with HGC-27 and GES-1 cells(F=29.503,31.153,87.750;all P＜0.05).Website prediction and dual luciferase experiments showed that LINC00461,MYH9,and miR-1301-3p had a targeting relationship.The relative luciferase activity of LINC00461-WT+miR-1301-3p mimic group was lower than that of LINC00461-WT+miR-NC group(t=9.202,P＜0.05).The relative luciferase activity of MYH9-WT+miR-1301-3p mimic group was lower than that of MYH9-WT+miR-NC group(t=10.944,P＜0.05).Compared with the si-NC group,the levels of LINC00461 and MYH9 mRNA,the positive rate of Edu and the expressions of CyclinD1 and Ki67 were decreased in si-LINC00461 group(t=7.351,9.992,15.424,9.640,11.192),while the levels of miR-1301-3p,apoptosis rate and the expressions of Bax and Caspase-3 were increased(t=8.104,34.304,6.341,7.473)(all P＜0.05).Compared with si-LINC00461+anti-NC group,the levels of MYH9 mRNA,the positive rate of Edu and the expressions of CyclinD1 and Ki67 were increased in si-LINC00461+anti-miR-1301-3p group(t=8.568,13.630,7.851,8.828),the level of miR-1301-3p,apoptosis rate and expression of Bax and Caspase-3 were decreased(t=6.549,22.847,4.726,4.941)(all P＜0.05).Conclusion Silencing LINC00461 could inhibit the proliferation of gastric cancer cells,promote cell apoptosis,and reduce cisplatin resistance,and the mechanism might be related to the regulation of miR-1301-3p/MYH9 axis.
Long non coding RNA LINC00461Micro RNA-1301-3pMyosin heavy chain 9Gastric cancerProliferation
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