摘要
目的探讨LIN9基因在人宫颈癌细胞株CasKi和HeLa中的表达及其与人乳头瘤病毒(HPV)E7表达的关系。方法利用qRT-PCR技术检测LIN9基因在宫颈癌细胞株CasKi(HPV16+)和HeLa(HPV18+)中的表达水平;在人永生化表皮细胞株HaCaT(HPV-)中转染质粒过表达HPV16/18E7、在人宫颈癌细胞株Caski和HeLa中转染siRNA沉默HPV16E7/18E7的表达,利用Western blot蛋白质印记法验证转染效果,应用q RT-PCR分别检测过表达及沉默E7表达后Ha Ca T、CasKi和He La三组细胞株中LIN9 mRNA的表达水平。结果 CasKi和HeLa细胞中LIN9的表达显著高于HaCaT细胞(1±0.22)(Cas Ki:2.55±0.25,t=22.63、P=0.026;He La:2.38±0.17,t=21.21、P=0.031)。Ha Ca T细胞过表达HPV16/18E7时,LIN9 mRNA表达水平亦显著高于对照组(1±0.14)(HPV16E7:1.65±0.16,t=13.12、P=0.036;HPV18E7:1.61±0.08,t=11.21、P=0.039);Cas Ki和He La细胞沉默HPV16E7/18E7表达时,LIN9 mRNA表达水平siRNA-NS组[(1±0.07)和(1±0.03)]均显著下降(HPV16E7:0.50±0.05,t=12.73、P=0.032;HPV18E7:0.57±0.11,t=14.30、P=0.035)。结论 LIN9在宫颈癌细胞中呈现高表达,且表达水平与E7表达有关。LIN9可能是抑制E7诱发宫颈癌变的重要抑癌因子,有望成为监测宫颈细胞HPV早期感染及宫颈癌治疗效果的重要生物学标志物。
Objective To investigate the expression of LIN9 gene in human cervical cancer cell lines of CasKi and HeLa and its relationship with E7 expression of HPV.Methods The expressions of LIN9 gene in cervical cancer cell lines CasKi(HPV16+)and HeLa(HPV18+)were detected by qRT-PCR method.Using transfection plasmid to overexpress HPV16/18E7 in human immortalized epidermal cell line HaCaT(HPV-),and siRNA to silence HPV16E7/18E7 expression in cell lines of CasKi and HeLa.The transfection effect was verified by Western blot,and LIN9 mRNA expression levels in cell lines of HaCaT,Caski and HeLa after silent E7 expression were detected by qRT-PCR.Results The expressions of LIN9 in CasKi and HeLa cell lines were significantly higher than that in HaCaT cells(1±0.22)(LIN9 in CasKi:2.55±0.25;t=22.63,P=0.026.LIN9 in HeLa:2.38±0.17;t=21.21,P=0.031)and the expression of LIN9 mRNA was increased significantly than that of control group(1±0.14)when overexpressed HPV16/18E7 in HaCaT cells(HPV16E7:1.65±0.16;t=13.12,P=0.036.HPV18E7:1.61±0.08,t=11.21,P=0.039),while decreased significantly after silencing HPV16E7/18E7 expression in CasKi and HeLa cells[(1±0.07)and(1±0.03)](HPV16E7:0.50±0.05;t=12.73,P=0.032.HPV18E7:0.57±0.11;t=14.30,P=0.035).Conclusions LIN9 was highly expressed in cervical cancer cells,and the expression level was related to the expression of E7.LIN9 may become an important biological marker for monitoring the early HPV infection of cervical cells and the therapeutic effect of cervical cancer.
作者
王鲲鹏
袁逸杰
朱嘉琪
张婷婷
Wang Kunpeng;Yuan Yijie;Zhu Jiaqi;Zhang Tingting(Department of Oral and Maxillofacial Surgery,Stomatological Hospital of Tianjin Medical University,Tianjin 300070,China)
出处
《中华实验和临床感染病杂志（电子版）》
CAS
2021年第3期196-201,共6页
Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)
基金
天津市教委科研计划项目(No.2018KJ053)
全国医学专业学位研究生教育指导委员会研究课题(No.B2-YX20190302-11)。
关键词
乳头状瘤病毒感染
宫颈肿瘤
生物标志物
肿瘤
Papillomavirus infection
Uterine cervical neoplasms
Biomarkers,tumor
