CircNRIP1调节miR-136-5p/RAC1轴对乳腺癌细胞增殖、凋亡和化疗耐药性的影响OACSTPCD
Effects of CircNRIP1 on proliferation,apoptosis and chemotherapy resistance of breast cancer cells through regulating miR-136-5p/RAC1 axis
目的 探讨CircNRIP1调节miR-136-5p/Ras相关C3肉毒素底物1(RAC1)轴对乳腺癌细胞增殖、凋亡和化疗耐药性的影响.方法 qRT-PCR法检测正常乳腺上皮细胞MCF10A、乳腺癌MCF-7细胞和紫杉醇(PTX)耐药细胞株MCF-7/PTX中CircNRIP1、miR-136-5p、RAC1 mRNA表达.将MCF-7/PTX细胞分为CK组(正常培养)、si-NC组(转染si-NC)、si-CircNRIP1组(转染si-CircNRIP1)、si-CircNRIP1+inhibitor NC 组(转染 si-CircNRIP1 和 inhibitor NC)、si-CircNRIP1+miR-136-5p inhibitor 组(转染si-CircNRIP1和miR-136-5p inhibitor).CCK-8法检测各组细胞增殖率;流式细胞术检测各组细胞凋亡情况;qRT-PCR法检测各组细胞CircNRIP1、miR-136-5p、RAC1 mRNA表达;Western blot检测各组细胞Ki-67、Bax、Bcl-2、RAC1蛋白表达量;双荧光素酶实验验证miR-136-5p与CircNRIP1、RAC1的靶向关系.结果 与正常乳腺上皮细胞MCF10A相比,MCF-7和MCF-7/PTX细胞中CircNRIP1、RAC1的表达升高(P<0.05),而miR-136-5p的表达降低(P<0.05);与MCF-7细胞相比,MCF-7/PTX细胞中CircNRIP1、RAC1的表达升高(P<0.05),miR-136-5p的表达降低(P<0.05).与CK组和si-NC组相比,si-CircNRIP1组MCF-7/PTX细胞增殖率,CircNRIP1和RAC1 mRNA表达,Bcl-2、Ki-67、RAC1蛋白表达降低(P<0.05),细胞凋亡率、miR-136-5p和Bax表达升高(P<0.05).敲低miR-136-5p表达可减弱沉默CircNRIP1对MCF-7/PTX细胞的抑制作用(P<0.05).双荧光素酶实验证实miR-136-5p与CircNRIP1和RAC1存在靶向关系.结论 沉默CircNRIP1表达可抑制MCF-7/PTX细胞恶性生物学行为,降低其PTX耐药性,可能与调控miR-136-5p/RAC1轴有关.
Objective To investigate the effects of CircNRIP1 on the proliferation,apoptosis and chemotherapy resistance of breast cancer cells by regulating miR-136-5p/Ras related C3 botulinum toxin substrate 1(RAC1)axis.Methods The mRNA expression of CircNRIP1,miR-136-5p and RAC1 in normal breast epithelial cells of MCF10A,breast cancer cells of MCF-7 and paclitaxel(PTX)resistant cell line of MCF-7/PTX were detected by qRT-PCR.MCF-7/PTX cells were divided into the CK group(normal culture),the si-NC group(transfected with si-NC),the si-CircNRIP1 group(transfected with si-CircNRIP1),the si-CircNRIP1+inhibitor NC group(transfected with si-CircNRIP1 and inhibitor NC),and the si-CircNRIP1+miR-136-5p inhibitor group(transfected with si-CircNRIP1 and miR-136-5p inhibitor).The cell proliferation rate of each group was detected by CCK-8 method;the cell apoptosis of each group was detected by flow cytometry;the expression of CircNRIP1,miR-136-5p,and RAC1 mRNA of each group were detected by qRT-PCR;the expression of Ki-67,Bax,Bcl-2,and RAC1 proteins of each group were detected by Western blot;the relationships between miR-136-5p and CircNRIP1 and RAC1 were verified by dual luciferase experiment.Results Compared with the normal breast epithelial cells of MCF10A,the expression of CircNRIP1 and RAC1 in the MCF-7 and MCF-7/PTX cells were increased(P<0.05),the expression of miR-136-5p was decreased(P<0.05);compared with the MCF-7 cells,the expression of CircNRIP1 and RAC1 in the MCF-7/PTX cells were increased(P<0.05),while the expression of miR-136-5p was decreased(P<0.05).Compared with the CK group and the si-NC group,the cell proliferation rate,the expression of CircNRIP1 and RAC1 mRNA,and the protein expression of Ki-67,Bcl-2,and RAC1 in the si-CircNRIP1 group were decreased(P<0.05),the apoptosis rate,and the expression of miR-136-5p and Bax were increased(P<0.05).Knocking down the expression of miR-136-5p could weaken the inhibitory effect of silencing CircNRIP1 on MCF-7/PTX cells(P<0.05).The dual luciferase experiment verified that miR-136-5p had targeting relationships with CircNRIP1 and RAC1.Conclusion Silencing CircNRIP1 expression can inhibit the malignant biological behavior of MCF-7/PTX cells,and reduce their PTX resistance,which may be related to regulating the miR-136-5p/RAC1 axis.
董路;李明;徐建立;许艳华
沧州市人民医院甲乳外科,河北 沧州 061000
临床医学
CircNRIP1miR-136-5p/RAC1轴乳腺癌化疗耐药紫杉醇
CircNRIP1miR-136-5p/RAC1 axisbreast cancerchemotherapy resistancepaclitaxel
《局解手术学杂志》 2024 (005)
383-387 / 5
河北省卫生健康委办公室医学科学研究课题计划(20232094)
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