人肿瘤转移抑制基因NM23-H1抑制卵巢癌转移机制体内体外实验研究

目的研究卵巢癌的人转移抑制基因NM23H1转移抑制的机制。方法首先构建由人肿瘤转移抑制基因NM23H1及真核细胞表达载体PcDNA3.1/zero构建成重组质粒PcDNA.NM23H1,并将其转染到高转移卵巢癌细胞株HO8910。体外实验:通过细胞DNA合成的流式细胞仪分析,以发现NM23H1对癌细胞生长的改变采用成集落实验,以观察转染组对转化生长因子(TGF)β刺激形成的集落数及细胞数并与对照组对比。同时检测转染组对化疗药物顺铂的敏感性。体内实验:将转染细胞注射于裸鼠皮下观察成瘤时间和肿瘤转移情况。结果NM23H1对卵巢癌细胞生长有一定抑制作用;实验组与对照组比较,实验组所形成的集落少,集落中细胞数目也少;被转染的细胞比对照组对顺铂更敏感,细胞死亡数多;裸鼠体内实验表明NM23H1对卵巢癌有抑制生长作用,主要对淋巴道转移有一定程度的抑制作用。结论NM23H1经高转移卵巢癌细胞株的体外实验发现对卵巢癌细胞生长及转移均有一定抑制作用;可增加对顺铂治疗的敏感性;体内实验表明,NM23H1可抑制癌瘤生长,对卵巢癌淋巴道转移有一定抑制作用。

Study of the mechanism of human metastasis suppressor gene NM23-H1 in suppressing the metastasis of ovarian carcinoma by using in-vitro and in-vivo experiments

Objective: To study the mechanism of human metastasis suppressor gene NM23-H1 in suppressing the metastasis of ovarian carcinoma.Methods: The recombinant PcDNA.NM23-Hl was constructed with mammalian expression vector Pc- DNA3.1/zero and human metastasis suppressor gene NM23-H1. The cultured human ovarian carcinoma cells (HO-8910 cell line) were transfected by the recombinant Pc-DNA.NM23-H1. We observed the cell growth, cell colonization, DNA contents in the cells, cell sensitivity to cisplatin and cell apoptosis by means of cell mitosis index curve, colony-forming test, flow cytometry, cell growth curve and DNA ladder methods. In-vivo experiments were performed by injecting the transfected cells hypocutaneously along the back of the nude mice to observe the rate of tumor growth and metastasis.Results: The results indicated that NM23-H1 gene had the effects of inhibition of the cell growth. The experimental group, compared with the control group, showed less cell mitosis and less colony formation. The transfected cells showed high sensitivity to cisplatin. In-vivo test showed slower tumor growth and less metastasis of the transfected cells.Conclusions: Our results suggested that NM23-H1 gene had a definite suppressive effect on the ovarian cancer growth and metastasis and also the effect of increasing the sensitivity of tumor cells to cisplatin. In-vivo nude mice experienments showed that NM23-H1 gene had the effect of suppressing the growth and metastasis of ovarian carcinoma, especially for the lymphatic metastasis.