miR-21对唾液腺腺样囊性癌细胞增殖和凋亡的影响及机制研究

doi:10.19438/j.cjoms.2017.03.004

中国口腔颌面外科杂志 ›› 2017, Vol. 15 ›› Issue (3): 209-213.doi: 10.19438/j.cjoms.2017.03.004

miR-21对唾液腺腺样囊性癌细胞增殖和凋亡的影响及机制研究

严斐¹，王超¹，黎婷¹，蔡文燕²，孙晋虎¹，²

1.徐州医科大学口腔医学院，江苏徐州 221000；
2.南京医科大学附属儿童医院口腔科，江苏南京 210000；
3.徐州医科大学附属医院口腔科，江苏徐州 221000

收稿日期:2016-10-11 修回日期:2017-01-12 出版日期:2017-05-20 发布日期:2017-06-09
通讯作者: 江苏省高校“青蓝工程”中青年学术带头人资助项目（51934）；徐州医学院优秀人才基金资助项目（2013012）
作者简介:严斐（1991-），女，硕士，E-mail:yffeifei@126.com
基金资助:
孙晋虎，E-mail: kqxsjh@163.com

Effect and underlying mechanism of miR-21 on proliferation and apoptosis of salivary adenoid cystic carcinoma cells

YAN Fei¹, WANG Chao¹, LI Ting¹, CAI Wen-yan², SUN Jin-hu¹，².

1.Department of Oral Medicine, School of Stomatology, Xuzhou Medical University. Xuzhou 221000；
2.Department of Stomatology, Children's Hospital Affiliated to Nanjing Medical University. Nanjing 210000；
3.Department of Stomatology, Affiliated Hospital of Xuzhou Medical University. Xuzhou 221004, Jiangsu Province, China

Received:2016-10-11 Revised:2017-01-12 Online:2017-05-20 Published:2017-06-09

摘要/Abstract

摘要： 目的：探讨miR-21对人唾液腺腺样囊性癌细胞株(SACC-LM)增殖、凋亡的影响及其相关作用机制。方法：将SACC-LM细胞分为 3 组，采用LipofectamineTM2000做瞬时转染。干扰组转染miR-21 inhibitor，阴性对照组转染无关核苷酸序列，空白对照组不转染。采用实时荧光定量PCR检测转染后细胞中miR-21和PDCD4、Bcl-2 mRNA的表达水平。以CCK8法检测转染24、48、72、96 h后细胞的增殖活性，Annexin-V/PI 双染色流式细胞术检测细胞凋亡率，Western免疫印迹检测转染后靶蛋白 PDCD4、Bcl-2的表达量。采用 SPSS 16.0软件包对所得数据进行统计学分析。结果：转染miR-21 inhibitor的细胞中miR-21、Bcl-2表达下调（P＜0.01），而PDCD4基因表达上调（P<0.05）。CCK8检测结果表明，miR-21表达下调可以明显抑制SACC-LM细胞的增殖活性，并呈时间依赖性（P<0.05）。流式细胞术检测结果表明，干扰组细胞凋亡率显著高于阴性对照组和空白组（P<0.05）；Western 免疫印迹结果显示， PDCD4蛋白的表达较对照组显著升高（P<0.01），Bcl-2蛋白表达显著降低（P<0.01）。结论：下调miR-21的表达可抑制SACC-LM细胞的增殖活性，诱导细胞凋亡，其机制可能与细胞内PDCD4表达上调，Bcl-2表达下调有关。

关键词: microRNA-21, 唾液腺腺样囊性癌, 增殖, 凋亡, PDCD4, Bcl-2

Abstract: PURPOSE: To investigate the effect and regulatory mechanisms of miR-21 on proliferation and apoptosis of human salivary adenoid cystic carcinoma (SACC-LM) cells. METHODS: SACC-LM cells were transfected with miR-21 inhibitor by LipofectamineTM 2000. Meanwhile, SACC-LM cells were transfected with NC inhibitor as negative control. Cell survival rate was calculated with CCK-8 assay at 24, 48, 72 and 96 h after transfection. The apoptosis rate was measured by flow cytometry. qRT-PCR analysis and Western blot were used to detect the expression of miR-21, PDCD4 and Bcl-2 in SACC-LM cell lines after transfection. SPSS 16.0 software package was used for statistical analysis. RESULTS: miR-21 expression was remarkably downregulated in miR-21 inhibitor-transfected cells. In the meantime, gene expression of PDCD4 was increased and Bcl-2 was decreased (P<0.05). miR-21 silencing triggered to a decreased cell viability in a time-dependent manner compared with the control groups (P<0.05). The apoptosis rate was dramatically increased compared with the control groups (P<0.01), and PDCD4 protein expression was significantly increased (P<0.05), while Bcl-2 was significantly decreased (P<0.05). CONCLUSIONS: Downregulation of miR-21 can inhibit proliferation of SACC-LM cells and induce apoptosis. The mechanism may be related to the increased expression of PDCD4 and decreased expression of Bcl-2.

Key words: miR-21, Salivary adenoid cystic carcinoma, Proliferation, Apoptosis, PDCD4, Bcl-2

中图分类号:

R739.8

严斐，王超，黎婷，蔡文燕，孙晋虎，. miR-21对唾液腺腺样囊性癌细胞增殖和凋亡的影响及机制研究[J]. 中国口腔颌面外科杂志, 2017, 15(3): 209-213.

YAN Fei, WANG Chao, LI Ting, CAI Wen-yan, SUN Jin-hu，.. Effect and underlying mechanism of miR-21 on proliferation and apoptosis of salivary adenoid cystic carcinoma cells[J]. China J Oral Maxillofac Surg, 2017, 15(3): 209-213.

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miR-21对唾液腺腺样囊性癌细胞增殖和凋亡的影响及机制研究

Effect and underlying mechanism of miR-21 on proliferation and apoptosis of salivary adenoid cystic carcinoma cells

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