Construction of eukaryotic expression vector carrying human TSLC1 gene and its expression in HepG2 cells

Objective： To construct the eukaryotic expression vector for human TSLC1 gene, and to express TSLC1 in HepG2 cells for investigating its effect on HepG2 cell growth. Methods： Full length of TSLC1 cDNA was amplified from RNA of normal human liver by RT-PCR, and cloned into pCI-neo expression vector. The recombinant plasmid pCI-TSLC1 was identified with restriction enzyme and sequenced, and then was stably transfected into HepG2 cells with lipofectamine 2000. The positive clones were examined by western-blotting and immunofluorescence, cell growth was analyzed with MTT assay. Results： The eukaryotic expression vector pCI-TSLC1 was successfully constructed and the stable cell line highly expressing TSLC1 protein was obtained. The growth of TSLCl-transfected cells was significantly suppressed in vitro. Conclusion： The HepG2 stable cell line could highly express TSLC1 protein, which provided a basis for further exploring the roles of TSLC1 in hepatocellular carcinoma.

2个基因在宫颈不同病变表达、与HPV感染相关性及诊断宫颈癌的意义

目的研究脆性组氨酸三联体基因(FHIT)及肺癌抑制因子(TSLC1)基因在宫颈不同病变的表达、与人乳头瘤(HPV)感染相关性及诊断宫颈癌的意义。方法选取2016年6月-2017年6月纳入182例妇科患者的临床资料,根据诊断结果分为:对照组(正常人)45例、宫颈内上皮瘤变(CIN)Ⅰ组45例、宫颈内上皮瘤变(CIN)组Ⅱ-Ⅲ55例和宫颈癌组37例。观察TSLC1和FHIT在正常宫颈、CIN及宫颈癌组织中的基因表达情况,研究TSLC1与FHIT在宫颈癌临床病理中的特点,分析2种基因与HPV感染的相关性,探讨TSLC1与FHIT在诊断宫颈癌的价值。结果 4组的高危HPV感染率有差异(P<0.01),4组的FHIT和TSLC1基因表达有差异(P<0.01)。FHIT、TSLC1表达在不同FIGO分期、不同分化及病理类型无统计学差异(P>0.05)。CINⅠ-Ⅲ组与宫颈癌组患者FHIT、TSLC1基因表达正常时的HPV感染率为0例(0.00%),FHIT、TSLC1基因表达缺失的HPV感染率升高(P<0.05)。FHIT、TSLC1基因表达情况与HPV感染呈显著性相关,两种基因表达缺失的患者HPV感染率升高(P<0.05)。FHIT基因缺失诊断宫颈癌的ROC的灵敏性为85.64%,特异性为82.33%;FHIT基因缺失诊断宫颈癌的ROC的灵敏性为88.52%,特异性为83.49%;两者缺失联合诊断宫颈癌的ROC的灵敏性为92.46%,特异性为95.32%,FHIT联合TSLC1基因缺失诊断宫颈癌的临床价值较单独基因缺失诊断价值高,差异显著(P<0.05)。结论TSLC1、FHIT基因表达在宫颈癌和HPV发生早期时会出现表达缺失。高危HPV感染患者早期行TSLC1、FHIT基因检测可提高宫颈癌的诊断率,此方法值得临床推广应用。