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目的研究miR-101对乳腺癌细胞增殖和迁移能力的影响,并分析miR-101影响乳腺癌细胞生物学行为的可能机制。方法采用实时荧光定量PCR检测miR-101和DNA甲基转移酶3a(DNMT3a)在乳腺癌组织和相对应的正常乳腺组织,正常乳腺细胞以及多种乳腺癌细胞系中的表达水平,应用慢病毒介导感染的方法分别将miR-101序列及shRNA-DNMT3a转至MDAMB-231人乳腺癌细胞,通过Western blot法检测DNMT3a以及上皮钙黏素(E-cadherin)的表达,分析miR-101影响乳腺癌细胞生物学行为的可能机制。通过MTT法检测乳腺癌细胞的增殖能力,划痕实验检测乳腺癌细胞的迁移能力。结果miR-101在MCF-7、MDA-MB-231、MDA-MB-361、HCC70乳腺癌细胞系及乳腺癌组织中表达低于正常乳腺组织及MCF-10A乳腺细胞系,而DNMT3a表达则升高。Western blot实验结果显示过表达miR-101的MDA-MB-231细胞中,DNMT3a的表达明显下降,而E-cadherin的表达升高;应用shRNA-DNMT3a抑制DNMT3a表达后,E-cadherin的表达也明显升高,证实miR-101可能通过抑制DNMT3a而发挥作用。体外实验显示过表达miR-101可明显抑制乳腺癌细胞MDA-MB-231的增殖和迁移能力。结论miR-101抑制乳腺癌细胞增殖和迁移能力可能是通过抑制DNMT3a进而恢复E-cadherin表达来实现。
Objective To investigate the effect of miR-101 on the proliferation and migration of breast cancer cells and to analyze the possible mechanism of miR-101 on the biological behavior of breast cancer cells. Methods The expression levels of miR-101 and DNMT3a in breast cancer tissues, corresponding normal breast tissues, normal breast cells and various breast cancer cell lines were detected by real-time fluorescence quantitative PCR. The lentivirus Mediated miR-101 and shRNA-DNMT3a were transfected into MDAMB-231 human breast cancer cells. The expression of DNMT3a and E-cadherin was detected by Western blot. The effect of miR-101 on breast cancer Possible mechanism of cancer cell biological behavior. The proliferation of breast cancer cells was detected by MTT assay. The scratch test was used to detect the migration ability of breast cancer cells. Results The expression of miR-101 in MCF-7, MDA-MB-231, MDA-MB-361 and HCC70 breast cancer cell lines and breast cancer tissues was lower than that in normal breast tissues and MCF- high. The results of Western blot showed that the expression of DNMT3a was significantly decreased and the expression of E-cadherin was increased in MDA-MB-231 cells overexpressing miR-101. The expression of E-cadherin was also significantly inhibited by DNMT3a shRNA-DNMT3a Increased, confirming that miR-101 may play a role by inhibiting DNMT3a. In vitro experiments showed that overexpression of miR-101 significantly inhibited the proliferation and migration of breast cancer cells MDA-MB-231. Conclusion The inhibitory effect of miR-101 on the proliferation and migration of breast cancer cells may be through the inhibition of DNMT3a and the restoration of E-cadherin expression.