口腔医学研究

口腔医学研究 ›› 2018, Vol. 34 ›› Issue (2): 152-156.DOI: 10.13701/j.cnki.kqyxyj.2018.02.011

• 口腔肿瘤学研究 • 上一篇    下一篇

RNA干扰沉默钙蛋白酶1对口腔鳞状细胞癌细胞株HSC3增殖和迁移的影响

张厚华1*, 范华俐2   

  1. 1. 荆州市中医医院口腔科 湖北 荆州 434000;
    2. 长沙市口腔医院 湖南 长沙 410000;
  • 收稿日期:2017-10-09 出版日期:2018-02-28 发布日期:2018-02-26
  • 通讯作者: 张厚华,E-mail:zhanghouhua1972@sina.com
  • 作者简介:张厚华(1972~ ),男,湖北武汉人,硕士,主治医师,主要从事口腔科疾病的基础与临床研究。
  • 基金资助:
    湖北省自然科学基金资助项目(编号:2015CFB089)

Effect of Calpain 1 Gene Silencing by RNA Interference on Proliferation and Migration of Oral Squamous Cell Carcinoma Cell Lines HSC3

ZHANG Hou-hua1*, FAN Hua-li2   

  1. 1. Department of Stomatology, Jingzhou Hospital of Traditional Chinese Medicine, Jingzhou 434000, China;
    2. Changsha Stomatological Hospital, Changsha 410000, China.
  • Received:2017-10-09 Online:2018-02-28 Published:2018-02-26

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摘要： 目的:探究RNA干扰使钙蛋白酶1(calpain 1)沉默后,对口腔鳞状细胞癌(OSCC)细胞株HSC3增殖和迁移的影响。方法:将calpain 1基因的siRNA表达载体(siRNA-calpain 1),瞬时转染至HSC3中。应用MTT法比较转染后细胞增殖情况;应用流式细胞术检测细胞周期和凋亡情况;应用Transwell测定评估细胞迁移和侵袭能力;应用蛋白免疫印迹检测细胞内TGF-β RII、Vimentin、Smad2、Smad3、磷酸化Smad2/3、E-cadherin、N-cadherin、MMP-2、MMP-9蛋白表达。结果:与NOKSI细胞系相比,HSC3细胞系中calpain 1表达显著增加(P＜0.05)。与Blank组和si-NC组相比,si-calpain 1 组calpain 1 表达均显著降低(P＜0.05)。与si-NC组相比,在转染si-calpain1后48 h、76 h和96 h,si-calpain 1组细胞活力均显著降低(P＜0.05);si-calpain 1组G0/G1期细胞比例显著增加,S期细胞比例显著降低(P＜0.05);si-calpain 1组细胞内E-cadherin表达显著增加,而cyclin-D1、cyclin-E、Vimentin、N-cadherin、MMP-2、MMP-9、TGF-β RII、Smad2、Smad3、p-Smad2/3蛋白表达均显著降低(P＜0.05)。结论:siRNA沉默calpain 1基因能够通过使细胞周期G0/G1停滞而抑制细胞增殖,并通过抑制MMPs及EMT来抑制细胞迁移和侵袭,这一过程涉及TGF-β/Smad信号通路。

关键词: 口腔鳞状细胞癌, 钙蛋白酶1, 细胞株HSC3, 细胞增殖, 细胞迁移

Abstract: Objective: To explore the silencing effect of calpain 1 by RNA interference on cell proliferation and migration in oral squamous cell carcinoma cell line HSC3. Methods: siRNA expression vector (siRNA-calpain 1) against the calpain 1 gene was transiently transfected into HSC3. The proliferation was tested by MTT method. The cell cycle and apoptosis was detected by flow cytometry. The cell migration and invasion were assessed by Transwell assay. The expression of TGF-β RII, Vimentin, Smad2, Smad3, phosphorylated Smad2/3, E-cadherin, N-cadherin, MMP-2, and MMP-9 protein were detected by western blot. Results: The expression of calpain 1 in HSC3 cell lines was significantly higher than those in NOKSI cell lines (P<0.05). Compared with the blank group and the si-NC group, the expression of calpain 1 was significantly decreased (P<0.05). Compared with si-NC group, the viability of si-calpain group was significantly decreased after 48 h, 76 h and 96 h after transfection with si-calpain1 (P<0.05). The proportion of G0/G1 phase cells in si-calpain group was significantly increased, while the proportion of S phase cells was significantly decreased (P<0.05). The expression of E-cadherin in si-calpain group was significantly increased, while cyclin-D1, cyclin-E, Vimentin, N-cadherin, MMP-2, MMP-9, TGF-β RII, Smad2, Smad3, p-Smad2/3 protein expression were significantly decreased (P<0.05). Conclusion: siRNA silencing calpain 1 gene can inhibit cell proliferation by inhibiting cell cycle G0/G1, and inhibit cell migration and invasion by inhibiting MMPs and EMT. This process involves TGF-β/Smad signaling pathway.

Key words: Oral squamous cell carcinoma, Calpain 1, Cell line HSC3, Cell proliferation, Cell migration

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