microRNA-21对人甲状腺乳头状癌细胞增殖和凋亡的影响

目的:探讨microRNA-21(miR-21)在人甲状腺乳头状癌中的表达及其对人甲状腺乳头状癌K1细胞增殖和凋亡能力的影响?方法:通过qRT-PCR检测12对甲状腺乳头状癌组织(PTC)与癌旁正常组织中miR-21的表达差异;将miR-21抑制试剂(Anti-miR-21)?过表达试剂(miR-21 mimic)分别转染入K1细胞,并各自设立阴性对照组,运用qRT-PCR验证K1细胞中miR-21的表达;MTT实验检测miR-21对K1细胞增殖能力的影响;流式细胞术检测转染后K1细胞凋亡的变化,Western blot检测细胞凋亡增殖相关蛋白的表达情况?结果:miR-21在PTC组织中的表达明显高于癌旁正常甲状腺组织(P < 0.05);与阴性对照组相比,瞬时转染Anti-miR-21的K1细胞miR-21的表达明显减弱(P < 0.01);MTT实验结果显示抑制miR-21表达后,K1细胞的增殖能力明显下降(P < 0.05);流式细胞仪检测显示转染Anti-miR-21后的K1细胞凋亡明显增加,凋亡率为19.5%,阴性对照组的凋亡率为9.4%;Western blot结果显示转染Anti-miR-21组K1细胞Bcl-2表达降低,Bax表达升高?转染miR-21mimic的K1细胞增殖和凋亡无明显改动?结论:miR-21在人甲状腺乳头状癌中呈高表达,抑制miR-21的表达能显著降低K1细胞的增殖能力,增加K1细胞凋亡率?

Effect of microRNA-21 on proliferation and apoptosis of papillary thyroid carcinoma cells

Objective:To explore the expression of microRNA-21 (miR-21) in papillary thyroid carcinoma (PTC) and its effect on proliferation and apoptosis of K1 cell. Methods:Real-time quantitative PCR(qRT-PCR) was performed to detect and compare the expressions of miR-21 in 12 pairs of PTC tissues and adjacent normal tissues. K1 cell was respectively transfected with Anti-miR-21 and mimic-miR-21 and their corresponding negative control groups were constructed. The expression of miR-21 in K1 cells was identified by qRT - PCR. Cell proliferation after transfection with anti-miR-21 was analyzed using MTT assay. The cell apoptosis of K1 after transfection was analyzed by flow cytometry. Western blotting was performed to detect the expressions of proliferation and apoptosis-related proteins. Results:Compared with adjacent normal tissues,miR-21 in PTC tissues was significantly up-regulated (P < 0.05). Compared with the negative control (Anti-miR-NC) group,the miR-21 expression of K1 cells was significantly down-regulated after Anti-miR-21 transfection(P < 0.01). MTT assay showed that the inhibition of miR-21 expression significantly reduced the proliferation of K1 cells(P < 0.05). Flow cytometry showed that K1 apoptosis rate after Anti-miR-21 transfection was significantly increased to 19.5%,while the negative control (Anti-miR-NC) group was 9.4%. Western blotting showed that the expression of Bcl-2 was down-regulated and the expression of Bax was up-regulated in the anti-miR-21 treated K1 cells. Conclusion:The expression of miR-21 is up-regulated in PTC. Down-regulated expression of miR-21 significantly inhibits proliferation while promotes the apoptosis of K1 cell.