党参总皂苷通过调控miR-142-3p/MARCH7轴影响乳腺癌细胞MCF-7增殖和凋亡

Effects of the proliferation and apoptosis of breast cancer cell MCF-7 by total saponins from Codonopsis pilosula via the miR-142-3p/MARCH7 axis

目的研究党参总皂苷(TSC)对乳腺癌细胞MCF-7增殖和凋亡的影响及作用机制。方法乳腺癌细胞MCF-7分成对照、TSC处理、miR-NC、miR-142-3p、Anti-miR-NC、Anti-miR-142-3p、 si-NC、si-MARCH7、TSC+Anti-miR-NC、TSC+Anti-miR-142-3p、TSC+Anti-miR-142-3p+si-NC和TSC+Anti-miR-142-3p+si-MARCH7组,MTT法测定细胞增殖,流式细胞术测定细胞凋亡,qRT-PCR法检测miR-142-3p表达,Western blot检测MARCH7蛋白表达。生物信息学软件预测miR-142-3p的靶基因,荧光素酶报告系统鉴定二者的靶向关系。结果与对照组比较,TSC组乳腺癌细胞存活率降低,凋亡率升高,miR-142-3p表达量增加,MARCH7表达量降低(P<0.05)。与对照组和miR-NC组比较,miR-142-3p组乳腺癌细胞存活率降低,凋亡率升高,miR-142-3p表达量升高(P<0.05)。与对照组和si-NC组比,si-MARCH7组乳腺癌细胞存活率降低,凋亡率升高,MARCH7表达量降低(P<0.05)。与TSC+Anti-miR-NC组比较,TSC+Anti-miR-142-3p组乳腺癌细胞存活率升高,细胞凋亡率降低(P<0.05)。与TSC+Anti-miR-142-3p+si-NC组比较,TSC+Anti-miR-142-3p+si-MARCH7组乳腺癌细胞存活率降低,细胞凋亡率升高(P<0.05)。MiR-142-3p靶向负调控MARCH7。结论一定剂量的TSC对乳腺癌细胞MCF-7增殖具有一定的抑制作用以及凋亡促进作用,可能是通过调控miR-142-3p/MARCH7轴发挥作用。

Objective To study the effect of total saponins from Codonopsis pilosula(TSC) on the proliferation and apoptosis of breast cancer cell MCF-7 and the corresponding mechanism. Methods Breast cancer cells MCF-7 were divided into groups: control, TSC, miR-NC, miR-142-3 p, Anti-miR-NC、Anti-miR-142-3 p、 si-NC, si-MARCH7, TSC+Anti-miR-NC, TSC+Anti-miR-142-3 p, TSC+Anti-miR-142-3 p+si-NC and TSC+Anti-miR-142-3 p+si-MARCH7 group. Cell proliferation was measured by MTT method and cell apoptosis was measured by flow cytometry. The levels of miR-142-3 p were measured by qRT-PCR and the expressions of MARCH7 protein were detected by Western blot. The bioinformatics software was used to predict the target genes of miR-142-3 p and the luciferase reporter system was used to identify the target gene of miR-142-3 p. Results Compared with the control group, the survival rate of MCF-7 cells in the TSC group decreased, the apoptosis rate increased, the expression of miR-142-3 p increased, and the expression of MARCH7 decreased(P<0.05). Compared with the control and miR-NC groups, the survival rate of breast cancer cells in the miR-142-3 p group was lower, the apoptosis rate was higher, and the expression of miR-142-3 p was higher(P<0.05). Compared with the control and si-NC groups, the survival rate of breast cancer cells in the si-MARCH7 group was lower, the apoptosis rate was higher, and the expression of MARCH7 was lower(P<0.05). Compared with the TSC+Anti-miR-NC group, the breast cancer cell survival rate in the TSC+Anti-miR-142-3 p group increased, and the apoptosis rate decreased(P<0.05). Compared with the TSC+Anti-miR-142-3 p+si-NC group, the breast cancer cell survival rate in the TSC+Anti-miR-142-3 p+si-MARCH7 group decreased, and the apoptosis rate increased(P<0.05). MiR-142-3 p was downregulated in March7. Conclusion Codonopsis pilosula could inhibit the proliferation and promote the apoptosis of breast cancer cell MCF-7 by regulating the miR-142-3 p/MARCH7 axis.