miR-206通过钙网蛋白调控乳腺癌MDA-MB-231细胞凋亡

MiR-206 Regulation of Apoptosis of Breast Cancer MDA-MB-231 Cells Through Calreticulin

目的探讨miR-206通过钙网蛋白(calreticulin,CALR)调控乳腺癌MDA-MB-231细胞恶性生物学行为的影响及其机制。方法乳腺癌MDA-MB-231细胞培养培养完成后,MTT法检测乳腺癌MDA-MB-231细胞的增殖能力。将miR-206 mimics和/或Ad-CALR分别转染到乳腺癌MDA-MB-231细胞中,流式细胞术检测乳腺癌MDA-MB-231细胞的凋亡情况;Western blotting实验检测乳腺癌MDA-MB-231细胞内CALR、E-cadherin和N-cadherin的表达。结果作用48 h后,miR-206mimics组乳腺癌MDA-MB-231细胞内CALR的表达被明显抑制(P<0.05);miR-206 mimics+Ad-CALR组和Ad-CALR组乳腺癌MDA-MB-231细胞中CALR的表达则均明显提高(P<0.05),MTT检测结果显示细胞增殖抑制率为(52.96±3.12)%,与空白对照组比较有明显的统计学意义(P<0.05);miR-206 mimics+Ad-CALR和Ad-CALR作用乳腺癌MDA-MB-231细胞48 h后,细胞增殖抑制率分别为(26.64±1.78)%和(24.17±1.56)%,与空白对照组比较差异均有明显统计学意义(P<0.05)。miR-206 mimics作用MDA-MB-231细胞48 h后,细胞凋亡率为(21.96±1.72)%,与空白对照组比较差异有统计学意义(P<0.05);miR-206mimics+Ad-CALR和Ad-CALR作用乳腺癌MDA-MB-231干细胞48 h后,细胞凋亡率分别为(6.93±0.41)%和(7.12±0.67)%,与空白对照组比较差异均统计学意义(P<0.05)。与空白对照组比较,miR-206 mimics组乳腺癌MDA-MB-231细胞内E-cadherin的表达被增强,而N-cadherin的表达被抑制(P<0.05);miR-206 mimics+Ad-CALR组和Ad-CALR组乳腺癌MDA-MB-231细胞内E-cadherin的表达被明显抑制,而N-cadherin的表达被明显增强(P<0.05)。结论miR-206可以通过降低乳腺癌MDA-MB-231细胞中CALR的表达来抑制乳腺癌干细胞的恶性生物学行为。

Objective To investigate the effect and mechanism of miR-206 on the malignant biological behavior of breast cancer MDA-MB-231 cells through calreticulin(CALR).Methods After breast cancer MDA-MB-231 cells were cultured,the proliferation ability of MDA-MB-231 cells was detected by MTT assay.MiR-206 mimics and/or Ad-CALR were respectively transfected into breast cancer MDA-MB-231 cells.The apoptosis of breast cancer MDA-MB-231 cells was detected by flow cytometry.Western blotting assay was used to detect the expression of CALR,E-cadherin and N-cadherin in MDA-MB-231 cells.Results After 48 hours’treatment,the expression of CALR in MDA-MB-231 cells was significantly inhibited in miR-206mimics group(P<0.05),but remarkably increased in miR-206 mimics+Ad-CALR group and Ad-CALR group(P<0.05).MTT detection results showed that the cell proliferation inhibition rate was(52.96±3.12)%,with statistically significant difference from that in blank control group(P<0.05).After 48 hours’treatment of breast cancer MDA-MB-231 cells with miR-206 mimics+Ad-CALR,the cell proliferation inhibition rates were(26.64±1.78)%and(24.17±1.56)%respectively,and the differences were statistically significant compared with blank control group(P<0.05).After MDA-MB-231 cells were treated with miR-206 mimics for 48 hours,the apoptosis rate was(21.96±1.72)%,with statistically significant difference from that in blank control group(P<0.05).After breast cancer MDA-MB-231 stem cells were treated by miR-206mimics+Ad-CALR and Ad-CALR for 48 hours,the apoptosis rates were(6.93±0.41)%and(7.12±0.67)%respectively,and the differences were statistically significant compared with blank control group(P<0.05).Compared with blank control group,the expression of E-cadherin in breast cancer MDA-MB-231 cells in miR-206 mimics group was enhanced,while the expression of N-cadherin was inhibited(both P<0.05).The expression of E-cadherin in MDA-MB-231 cells in miR-206 mimics+Ad-CALR group and Ad-CALR group was significantly inhibited,while the expression of N-cadherin was significantly enhanced(both P<0.05).Conclusion MiR-206 can inhibit the malignant biological behavior of breast cancer stem cells by reducing the expression of CALR in breast cancer MDA-MB-231 cells.