miR-299-3p通过调控PAX3基因表达介导乳腺癌细胞增殖和凋亡  

miR-299-3p mediates the proliferation and apoptosis of breast cancer cells by regulating the expression of PAX3 gene

作　　者：陈飞 李甜 王建东[1] 刘晖 郑洁[1] 王凤 姜姝君 胡俊艳[1] 李喆 CHEN Fei;LI Tian;WANG Jian-dong;LIU Hui;ZHENG Jie;WANG Feng;JIANG Shu-jun;HU Jun-yan;LI Zhe(Department of Breast Surgery,Shuguang Hospital Affliated to Shanghai University Traditional Chinese Medicine,Shanghai 201203,China;Department of Breast Surgery,Baoshan Branch of Shuguang Hospital Affliated to Shanghai University Traditional Chinese Medicine,Shanghai 201999,China)

机构地区：[1]上海中医药大学附属曙光医院乳腺外科,上海201203 [2]上海中医药大学附属曙光医院宝山分院乳腺外科,上海201999

出　　处：《中国现代普通外科进展》2022年第1期1-6,共6页Chinese Journal of Current Advances in General Surgery

基　　金：上海市卫生和计划生育委员会中医药科研课题项目(2018LP036)。

摘　　要：目的:研究miR-299-3p对乳腺癌细胞增殖、凋亡的影响和潜在的分子机制。方法:以乳腺上皮细胞MCF-10A为对照,qRT-PCR检测乳腺癌MDA-MB-231和MDA-MB-468细胞中miR-299-3p和PAX3 mRNA的表达,Western blot检测MDA-MB-231和MDA-MB-468细胞中PAX3、增殖相关蛋白Ki-67、PCNA以及凋亡相关蛋白Bcl-2、Bax的表达,CCK-8法和流式细胞术分别测定MDA-MB-231和MDA-MB-468细胞增殖吸光度值和凋亡率,双荧光素酶报告系统验证miR-299-3p和PAX3的靶向关系。结果:与乳腺上皮细胞MCF-10A相比,在乳腺癌细胞MDA-MB-231、MDA-MB-468中miR-299-3p的含量显著减少(P<0.05),而PAX3 mRNA和蛋白表达量显著增加(P<0.05);过表达miR-299-3p可下调细胞吸光度值,提高细胞凋亡率,降低增殖凋亡相关蛋白Ki-67、PCNA和Bcl-2含量,上调Bax含量;miR-299-3p靶向负调控PAX3的表达;敲减PAX3可下调Ki-67、PCNA和Bcl-2蛋白含量,促进BAX表达,降低细胞吸光度值,提高细胞凋亡率;过表达PAX3逆转了miR-299-3p过表达对MDA-MB-231细胞增殖、凋亡的作用。结论:miR-299-3p通过靶向PAX3调控MDA-MB-231细胞增殖和凋亡,miR-299-3p可能是乳腺癌的潜在分子靶点。Objective: To investigate the effects of miR-299-3 p on proliferation and apoptosis of breast cancer cells and the potential mechanism. Methods: Taking the breast epithelial cells MCF-10 A as the control, the expression levels of miR-299-3 p and PAX3 mRNA in breast cancer MDA-MB-231 and MDA-MB-468 cells were measured by quantitative real-time polymerase chain reaction(q RT-PCR). The protein expression levels of PAX3, proliferation-related proteins Ki-67, proliferating cell nuclear antigen(PCNA), and apoptosis-related proteins BCl-2, Bax in MDA-MB-231 and MDA-MB-468 cells were determined by Western blot. Cell proliferation OD value and apoptotic rate were measured by CCK8 assay and flow cytometry, respectively. And the targeting relationship between miR-299-3 p and PAX3 was examined by dual-luciferase reporter assay system. Results:Compared with breast epithelial cells MCF-10 A, the levels of miR-299-3 p in breast cancer cells MDA-MB-231 and MDA-MB-468 were significantly reduced(P<0.05), while the expression of PAX3 mRNA and protein were significantly increased(P<0.05). Over-expression of miR-299-3 p down-regulated the OD value of MDA-MB-231 cells, increased the apoptosis rate, reduced the levels of Ki-67, PCNA and Bcl-2, and up-regulated the level of Bax proteins. Mi R-299-3 p targeted and negatively regulated the expression of PAX3. Knocking down PAX3 can down-regulated the levels of Ki-67, PCNA and Bcl-2 proteins, promoted the expression of Bax, reduced the OD value of MDA-MB-231 cells and increased the apoptosis rate. Over-expression of PAX3 reversed the effect of miR-299-3 p over-expression on proliferation and apoptosis of MDA-MB-231 cells. Conclusion: miR-299-3 p regulates the proliferation and apoptosis of breast cancer MDA-MB-231 cells by targeting PAX3. miR-299-3 p is a potential molecular target for breast cancer.

关 键 词：乳腺肿瘤 MDA-MB-231细胞 miR-299-3p PAX3 增殖 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]