目的:观察欣胃颗粒对胃癌细胞METTL16/CyclinD1通路和增殖、凋亡和细胞周期的影响，阐明其治疗胃癌的分子机制。方法:采用CCK8法检测不同浓度欣胃颗粒对胃癌细胞株(MKN-45)细胞活性的影响，采用流式细胞术检测细胞凋亡和细胞周期的分布，采用 Western blot和 RT-qPCR法检测METTL16/CyclinD1通路中相关分子的表达，采用斑点杂交检测CyclinD1的m6A甲基化修饰水平。结果:与空白组相比，欣胃颗粒能够显著促进MKN-45细胞凋亡水平，增加G0/G1 期比例，降低S期比例，下调CyclinD1、CDK4、E2F1和METTL16蛋白的表达和p-Rb/Rb的比值，差异均具有统计学意义(P<0。01)。此外，欣胃颗粒能够显著下调METTL16和CyclinD1 mRNA的表达，抑制CyclinD1的m6A甲基化修饰水平，差异均具有统计学意义(P<0。01)。结论:欣胃颗粒通过抑制METTL16介导的CyclinD1的m6A甲基化修饰水平，从而发挥阻断G1期向S期转化、抑制细胞增殖和促进细胞凋亡的作用。
Effects of Xinwei granules on the METTL16/CyclinD1 pathway in gastric cancer cell proliferation,apoptosis,and cell cycle
Objective:To investigate the effects of Xinwei granules on the METTL16/CyclinD1 pathway and proliferation,apoptosis,and cell cycle of gastric cancer cells,and to elucidate the molecular mechanisms underlying its therapeutic potential in gastric cancer.Methods:Cell viability of the MKN-45 gastric cancer cell line was assessed using the CCK-8 assay following treat-ment with various concentrations of gastric-derived granules.Flow cytometry was employed to analyze apoptosis and cell cycle dis-tribution.Western blot and RT-qPCR analyses were used to measure the expression of key molecules in the METTL16/CyclinD1 pathway.Spot hybridization was performed to detect m6A methylation modifications of CyclinD1.Results:Compared to the con-trol group,gastric-derived granules significantly increased apoptosis in MKN-45 cells,elevated the proportion of cells in the G1 phase,reduced the proportion in the S phase,and downregulated the expression of CyclinD1,CDK4,E2F1,and METTL16 proteins as well as the p-Rb/Rb ratio(P<0.01).Additionally,gastric-derived granules significantly decreased the mRNA expres-sion levels of METTL16 and CyclinD1,and inhibited CyclinD1 m6A methylation modifications(P<0.01).Conclusion:Xinwei granules exert their effects by inhibiting METTL16-mediated m6A methylation of CyclinD1,thereby blocking the transition from the G1 phase to the S phase,suppressing cell proliferation,and promoting apoptosis.
Xinwei granulesMETTL16/CyclinD1 signaling pathwaym6A methylation modificationsPrecancerous le-sions of gastric cancer
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