miR-140-5p对口腔鳞状细胞癌细胞增殖及凋亡的影响

›› 2017, Vol. 8 ›› Issue (4): 186-190.

miR-140-5p对口腔鳞状细胞癌细胞增殖及凋亡的影响

杜莉¹,曹伟靖¹,田莹¹,王原明²

1. 延安大学附属医院口腔科
2. 延安大学附属医院

收稿日期:2017-06-21 修回日期:2017-10-27 出版日期:2017-12-25 发布日期:2017-12-26
通讯作者: 王原明 E-mail:yuanmingkq@163.com

Effect of miR-140-5p on the proliferation and apoptosis of OSCC cells

Received:2017-06-21 Revised:2017-10-27 Online:2017-12-25 Published:2017-12-26

摘要/Abstract

摘要： 目的：研究miR-140-5p对口腔鳞状细胞癌（OSCC）增殖及凋亡的影响。方法：实时定量RT-PCR检测miR-140-5p在OSCC细胞系Tca8113、Cal27、SCC25中以及正常人口腔黏膜成纤维细胞系（hOMF）的表达；MTT法检测miR-140-5p mimic转染对Tca8113细胞增殖的影响；流式细胞仪检测miR-140-5p mimic转染对Tca8113细胞凋亡的影响；双荧光素酶报告基因检测miR-140-5p与微管解聚蛋白1（STMN1）的靶向关系；Western blot法检测hOMF和三种OSCC细胞中STMN1的表达，以及miR-140-5p mimic转染对Tca8113细胞中STMN1、音猬因子（Shh）、Smoothened（Smo）、Ptch、胶质母细胞瘤转录因子（Gli1）等的表达。结果：miR-140-5p在三种OSCC细胞系中的表达显著下调；过表达miR-140-5p可显著抑制Tca8113细胞增殖，促进细胞凋亡。双荧光素酶报告基因表明STMN1为miR-140-5p的靶向调节基因。STMN1在OSCC细胞中的表达显著上升，过表达miR-140-5p可显著抑制STMN1、Shh、Smo、Ptch、Gli1等的表达。结论：miR-140-5p可通过抑制STMN1的表达，抑制Hedgehog信号通路，来抑制OSCC细胞增殖，促进细胞凋亡。

Abstract: Objective：To analyze the effect of miR-140-5p on the proliferation and apoptosis of OSCC cells. Methods：The expression of miR-140-5p in Tca8113, Cal27, SCC25 cells and human oral mucosa fibroblasts (hOMF) was measured by Real-time PCR. The effect of miR-140-5p mimic on the proliferation of Tca8113 cells was measured by MTT method. The effect of miR-140-5p mimic on the cell apoptosis was measured by Flow cytometry. The target relationship between miR-140-5p and STMN1 was measured by dual luciferase reporter assay. Expressions of STMN1 in hOMF and three OSCC cells, and the effect of miR-140-5p mimic transfection on expression of Shh, Smo, Ptch and Gli1 were measured by western blot. Results：MiR-140-5p was down-regulated in OSCC cells. Overexpression of miR-140-5p suppressed Tca8113 cell proliferation and induced cell apoptosis. The result of dual luciferase reporter assay showed that STMN1 was the direct target of miR-140-5p. STMN1 was overexpressed in OSCC cells. Overexpression of miR-140-5p significantly reduced the expression of STMN1. Furthermore, overexpression of miR-140-5p decreased the expression of Shh, Smo, Ptch and Gli1. Conclusions：MiR-140-5p suppressed OSCC cell proliferation and induced cell apoptosis through down-regulating the expression of STMN1 and inhibiting the Hedgehog signaling pathway.

杜莉曹伟靖田莹王原明. miR-140-5p对口腔鳞状细胞癌细胞增殖及凋亡的影响[J]. , 2017, 8(4): 186-190.

参考文献

[1]李击, 王萍, 岑红兵, 等.miR-17-5p通过靶基因SOCS6促进口腔鳞癌细胞增殖的研究[J].临床口腔医学杂志, 2015, 31(11):656-659
[2]Liao L, Wang J, Ouyang S, et al.Expression and clinical significance of microRNA-1246 in human oral squamous cell carcinoma[J].Med Sci Monit, 2015, 21:776-781
[3]郭艳, 尚超, 任美思, 等.miR-15b对口腔鳞癌细胞Cal27增殖与凋亡能力影响研究[J].中国实用口腔科杂志, 2015, 8(8):458-460
[4]Chang L, Lei X, Qiu YU, et al.MicroRNA-133b inhibits cell migration and invasion by targeting matrix metalloproteinase 14 in glioblastoma[J].Onco Lett, 2015, 10(5):2781-2786
[5]Park H, Huang X, Lu C, et al.MicroRNA-146a and microRNA-146b regulate human dendritic cell apoptosis and cytokine production by targeting TRAF6 and IRAK1 proteins[J].J Biol Chem, 2015, 290(5):2831-2841
[6]Shao Y, Qu Y, Dang S, et al.MiR-145 inhibits oral squamous cell carcinoma (OSCC) cell growth by targeting c-Myc and Cdk6[J].Cancer Cell Int, 2013, 13(1):51-
[7]Wang XC, Ma Y, Meng PS, et al.miR-433 inhibits oral squamous cell carcinoma (OSCC) cell growth and metastasis by targeting HDAC6[J].Oral Oncol, 2015, 51(7):674-682
[8]Liu C, Wang Z, Wang Y, et al.MiR-338 suppresses the growth and metastasis of OSCC cells by targeting NRP1[J].Mol Cell Biochem, 2015, 398(1/2):115-122
[9]赵珍.microRNAs调控口腔鳞癌增殖及凋亡的实验研究 [D]. 郑州：郑州大学, 2015.
[10]Zhai H, Fesler A, Ba Y, et al.Inhibition of colorectal cancer stem cell survival and invasive potential by hsa-miR-140-5p mediated suppression of Smad2 and autophagy[J].Oncotarget, 2015, 6(23):19735-19746
[11]Lan H, Chen W, He G, et al.miR-140-5p inhibits ovarian cancer growth partially by repression of PDGFRA[J].Biomed Pharmacother, 2015, 75:117-122
[12]陈翔, 黄路, 张中卒, 等.miR-140在骨肉瘤组织中表达降低及其过表达可促进骨肉瘤U2细胞的凋亡[J].基础医学与临床, 2016, 36(4):439-444
[13]刘飞, 刘芳, 孙玉琳, 等.微管不稳定蛋白在食管癌细胞和组织中的表达及意义[J].世界华人消化杂志, 2010, 18(13):1306-1312
[14]Akhtar J.STMN1在食管鳞状细胞癌的临床和生物学[D].济南：山东大学, 2015.
[15]邱胜聪.miR-223-3p通过靶向作用STMN1调控胶质瘤细胞的生长和替莫唑胺化疗敏感性 [D].广州：南方医科大学, 2015.
[16]Li J, Kong F, Wu K, et al.miR-193b directly targets STMN1 and uPA genes and suppresses tumor growth and metastasis in pancreatic cancer[J].Mol Med Rep, 2014, 10(5):2613-2620
[17]严明, 王丽君, 陈万涛, 等.Sonic Hedgehog信号通路在口腔鳞癌干细胞中调控作用 [C]. 第十二次全国医学遗传学学术会议, 2013.
[18]张洪瑞, 武和明, 张萍, 等.口腔鳞癌中Shh/Gli1的表达及其临床生物学意义[J].中国口腔颌面外科杂志, 2010, 8(5):445-449
[19]Chung MK, Kim HJ, Lee YS, et al.Hedgehog signaling regulates proliferation of prostate cancer cells via stathmin1[J].Clin Exp Med, 2010, 10(1):51-57