摘要
目的探讨口腔癌细胞中SIRT7表达,分析其表达对口腔癌细胞EMT转化、转移能力的影响及可能分子机制。方法Western blotting检测SIRT7在正常口腔细胞系HOK和人口腔癌细胞系HSC3、OECM1、OC3、SCC4和SCC25中的表达;采用慢病毒载体过表达(OE-SIRT7组)或敲低(shSIRT7组)HSC3细胞系中的SIRT7表达;采用细胞划痕实验和Transwell实验检测SIRT7表达对口腔癌细胞迁移和侵袭的影响;Western blotting检测SIRT7、SMAD4、Ac-SMAD4、E-cadherin、N-cadherin、Vimentin和MMP-7蛋白表达。结果SIRT7在HSC3(0.43±0.032)、OECM1(0.27±0.043)、OC3(0.37±0.025)、SCC4(0.31±0.022)、SCC25(0.72±0.063)中的表达均低于HOK细胞的0.97±0.133。OE-SIRT7组细胞划痕愈合率[(13.54±4.35)]%和向下层小室侵袭率[(38.46±7.68)%]明显低于阴性对照组[(49.58±11.26)%、(60.14±15.74)%],而shSIRT7组细胞划痕愈合率[(76.58±17.26)%]和向下层小室侵袭率[(70.42±20.64)%]明显高于阴性对照组(P<0.05)。沉默SIRT7显著降低E-cadherin表达,并上调了Ac-SMAD4、N-cadherin、Vimentin和MMP-7蛋白水平;沉默SMAD4后,SIRT7敲降对Ac-SMAD4表达增加以及EMT相关蛋白分子的影响显著降低。结论SIRT7通过促进SMAD4去乙酰化抑制口腔癌细胞EMT转化,进而抑制口腔癌细胞侵袭和迁移。
Objective To investigate the expression of SIRT7 in oral cancer cells and analyze its effect on EMT transformation and metastasis and its molecular mechanism.Methods Western blotting was used to detect the expression of SIRT7 in HOK,HSC3,OECM1,OC3,SCC4 and SCC25.SIRT7 expression in HSC3 cell line was overexpressed(OE-SIRT7 group)or knockdown(shSIRT7 group)by lentivirus vector.The effects of SIRT7 expression on migration and invasion of oral cancer cells were detected by cell scratch test and Transwell test.Western blotting was used to detect the protein expression of SIRT7,SMAD4,Ac-SMAD4,E-cadherin,N-cadherin,Vimentin and MMP-7.Results Expression of SIRT7 in HSC3,OECM1,OC3,SCC4,SCC25 was significantly down-regulated compared with HOK cells.The wound healing ability[(13.54±4.35)%]and migration[(38.46±7.68)%]of HSC3 cells in OE-SIRT7 group were significantly lower than that of the negative control group[(49.58±11.26)%and(60.14±15.74)%],while the wound healing ability[(76.58±17.26)%]and migration[(70.42±20.64)%]of HSC3 cells in shSIRT7 group were significantly higher(P<0.001).SIRT7 silencing significantly reduced the expression of E-cadherin and up regulated the levels of Ac-SMAD4,N-cadherin,Vimentin and MMP-7.After SMAD4 silencing,SIRT7 knockdown significantly reduced the expression of Ac-SMAD4 and the effect of EMT related protein molecules.Conclusion SIRT7 can inhibit the EMT transformation of oral cancer cells by promoting the deacetylation of SMAD4,and then inhibit the invasion and migration of oral cancer cells.
作者
高运来
陶文
徐峰
李伟
GAO Yunlai;TAO Wen;XU Feng;LI Wei(Department of Stomatology,Jinan People’s Hospital,Jinan 271199,China)
出处
《临床肿瘤学杂志》
CAS
北大核心
2019年第12期1089-1093,共5页
Chinese Clinical Oncology
