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目的:探讨TIZ基因过表达对卵巢上皮癌细胞生物学特性的影响。方法:采用反转录(reverse transcription,RT)-PCR法从人卵巢癌组织总RNA中扩增TIZ基因的cDNA全长序列,与pcDNA3.1/myc-his(-)C质粒载体连接构建重组质粒;采用脂质体转染法将重组质粒pcDNA3.1/myc-his(-)C-TIZ(+)转染入卵巢上皮癌细胞HO8910中;分别采用MTT法和集落形成实验测定细胞的增殖能力;FCM法检测细胞周期;细胞体外侵袭重建基膜实验、Transwell小室实验和体外黏附实验分别测定细胞的体外侵袭、转移及黏附能力。结果:从卵巢癌组织中扩增获得了TIZ基因全长序列,并成功构建pcDNA3.1/myc-his(-)C-TIZ(+)重组质粒,测序证实插入片段序列正确。细胞生长曲线、细胞周期和细胞集落测定实验显示,pcDNA3.1/myc-his(-)C-TIZ(+)-HO8910细胞与对照组细胞相比较,其细胞增殖受到明显抑制,差异有统计学意义(P<0.05)。细胞体外侵袭、转移及黏附能力测定实验结果显示,HO8910细胞在TIZ基因过表达前后其侵袭、迁移和黏附能力无明显改变,差异无统计学意义(P>0.05)。结论:TIZ基因过表达对卵巢上皮癌细胞生长、增殖有一定的抑制作用,可能发挥抑癌基因的生物学作用。
Objective: To investigate the effect of TIZ gene overexpression on the biological characteristics of epithelial ovarian cancer cells. Methods: The full length cDNA of TIZ gene was amplified from total RNA of human ovarian cancer tissues by reverse transcription (RT) -PCR and ligated with pcDNA3.1 / myc-his (-) C plasmid vector to construct recombinant The recombinant plasmid pcDNA3.1 / myc-his (-) C-TIZ (+) was transfected into ovarian epithelial carcinoma cell line HO8910 by lipofectamine. The cell proliferation was measured by MTT assay and colony formation assay Ability; Cell cycle was detected by FCM; In vitro invasion and rebuilding basement membrane experiments, Transwell chamber experiments and in vitro adhesion assays were used to determine the invasion, metastasis and adhesion of cells in vitro. Results: The full length of TIZ gene was amplified from ovarian cancer tissue and the pcDNA3.1 / myc-his (-) C-TIZ (+) recombinant plasmid was successfully constructed. The sequence of the inserted fragment was confirmed by sequencing. Cell growth curve, cell cycle and colony assay showed that the cell proliferation of pcDNA3.1 / myc-his (-) C-TIZ (+) - HO8910 cells was significantly inhibited compared with the control cells Significance (P <0.05). The results of in vitro invasion, metastasis and adhesion assay showed that there was no significant difference in the invasion, migration and adhesion of HO8910 cells before and after TIZ gene overexpression (P> 0.05). Conclusion: Overexpression of TIZ may inhibit the growth and proliferation of epithelial ovarian cancer cells and may play a biological role in tumor suppressor genes.