过表达miR-138抑制TCF-4对甲状腺癌细胞CAL-62增殖，凋亡和运动的调节作用

doi:10.13418/j.issn.1001-165x.2019.04.013

中国临床解剖学杂志 ›› 2019, Vol. 37 ›› Issue (4) : 425-430. DOI: 10.13418/j.issn.1001-165x.2019.04.013

吴闽，　侯慧珍，　司廷林

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Regulatory effects of miR-138 overexpression on proliferation, apoptosis and movement of thyroid carcinoma cell line CAL-62 through inhibiting TCF-4

WU Min, HOU Hui-zhen, SI Ting-lin

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摘要

目的　探究过表达miR-138抑制T细胞因子4（TCF-4）对甲状腺癌细胞CAL-62增殖，凋亡和运动的调节作用机制。方法　转染miR-138 mimic于CAL-62细胞，使用RT-PCR检测miR-138的表达；使用TCF-4构建pcDNA过表达载体转染CAL-62细胞，使用Westen blot检测TCF-4、增殖核抗原67（Ki67）、增殖细胞核抗原（PCNA）、半胱天冬酶3（caspase-3）、半胱天冬酶9（caspase-9）、上皮型钙黏蛋白（E-cadherin）、波形蛋白（Vimentin）和靶基因cycD、c-Myc的表达；使用EDU染色检测细胞增殖；使用Hoechst染色检测细胞凋亡情况；transwell检测细胞侵袭能力。结果　相比空白组，miR-138转染+TCF-4组caspase-3、caspase-9、Vimentin蛋白表达显著减少，且有显著性差异（P<0.01）；Ki67、PCNA、E-cadherin、cycD、c-Myc蛋白表达显著增加（P<0.01）；甲状腺肿瘤细胞凋亡受到显著抑制（P<0.01），增殖、侵染受到显著的促进（P<0.01）。相比TCF-4组，miR-138转染+TCF-4组caspase-3、caspase-9、Vimentin蛋白表达显著增加、（P<0.01）；Ki67、PCNA、E-cadherin、cycD、c-Myc蛋白表达显著减少（P<0.01），抑制了甲状腺肿瘤细胞增殖、侵染（P＜0.01），凋亡受到了显著的促进（P<0.01）。结论　表达miR-138抑制TCF-4使得甲状腺癌细胞CAL-62的增殖、侵染受到显著抑制，凋亡受到显著促进。

Abstract

Objective　To explore the mechanism of regulatory effects of miR-138 overexpression through inhibiting TCF-4 expression on the proliferation, apoptosis and movement of thyroid carcinoma cell line CAL-62.　Methods　miR-138 mimic was transfected into CAL-62 cells, and detected by RT-PCR. After overexpression of pcDNA vector, TCF-4 was transfected into CAL-62 cells, and Western blot was used to detect the expression levels of TCF-4, proliferating nuclear antigen 67 (Ki67), proliferating cell nuclear antigen (PCNA), caspase-3, caspase-9, E-cadherin, Vimentin and target genes cycD and c-Myc. EDU staining was used to detect the cell proliferation. Hoechst staining was used to detect the cell apoptosis, and transwell was used to detect the cell invasion ability.　Results　Compared with the control group, the protein expression levels of caspase-3, caspase-9 and Vimentin in miR-138 transfection+TCF-4 group were significantly decreased (P<0.01). The protein expression levels of Ki67, PCNA, E-cadherin, cycD and c-Myc were significantly increased (P<0.01). And the apoptosis of thyroid tumor cells was significantly inhibited (P<0.01), and the proliferation and invasion were significantly promoted (P<0.01). Compared with TCF-4 group, the protein expression levels of caspase-3, caspase-9 and Vimentin in miR-138 transfection+TCF-4 group were significantly increased (P<0.01). The protein levels of Ki67, PCNA, E-cadherin, cycD and c-Myc were significantly decreased (P<0.01). Proliferation and invasion of thyroid tumor cells were significantly inhibited (P<0.01), as well apoptosis was significantly promoted (P<0.01).　Conclusions　miR-138 overexpression can significantly inhibit the proliferation and invasion and significantly promote the apoptosis of thyroid cancer cell line CAL-62, by inhibiting TCF-4 expression.

导出引用

吴闽, 侯慧珍, 司廷林. 过表达miR-138抑制TCF-4对甲状腺癌细胞CAL-62增殖，凋亡和运动的调节作用[J]. 中国临床解剖学杂志. 2019, 37(4): 425-430 https://doi.org/10.13418/j.issn.1001-165x.2019.04.013

WU Min, HOU Hui-zhen, SI Ting-lin. Regulatory effects of miR-138 overexpression on proliferation, apoptosis and movement of thyroid carcinoma cell line CAL-62 through inhibiting TCF-4[J]. Chinese Journal of Clinical Anatomy. 2019, 37(4): 425-430 https://doi.org/10.13418/j.issn.1001-165x.2019.04.013

中图分类号： R736.1

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