摘要
目的 NK4不仅具有肝细胞生长因子(HGF)拮抗剂的作用,而且具有抗血管生成作用。该实验通过在人卵巢癌细胞SKOV-3中转染NK4cDNA片段,研究NK4在裸鼠体内对人卵巢癌的治疗作用。方法 NK4和荧光霉素(luciferase)表达质粒分别转染人卵巢癌细胞SKOV-3,稳定转染后,得到SKOV-3/NK4细胞和对照(SKOV-3/LUC)细胞。检测NK4在细胞培养基中的表达情况以及经不同培养基(SKOV-3、SKOV-3/LUC和SKOV-3/NK4培养基上清)培养后SKOV-3细胞中c-Met和磷酸化-c-Met的表达,检测三组肿瘤细胞(SKOV-3、SKOV-3/LUC和SKOV-3/NK4)体外生长速度,建立裸鼠皮下肿瘤生长模型,比较三组肿瘤细胞在裸鼠体内的生长速度。结果在SKOV-3/NK4培养基中有NK4蛋白表达,对照(SKOV-3和SKOV-3/LUC)细胞中无表达。磷酸化-c-Met在SKOV-3/NK4培养基培养的SKOV-3细胞中被抑制,而在对照(SKOV-3和SKOV-3/LUC)细胞培养基培养的SKOV-3细胞中正常表达。c-Met表达在各组无差别。三种细胞体外生长曲线差异无显著性(P>0.05)。一周后SKOV-3/NK4细胞接种的肿瘤生长缓慢,SKOV-3和SKOV-3/LUC细胞接种的肿瘤生长迅速,与SKOV-3/NK4组相比有统计学意义(P<0.05)。结论 NK4蛋白在SKOV-3/NK4细胞培养基中大量分泌,NK4能抑制人卵巢癌细胞c-Met受体磷酸化。在裸鼠体内NK4能抑制卵巢癌细胞生长,NK4可作为卵巢癌基因治疗的新方法。
【Objective】 Hepatocyte growth factor（HGF） has its function after binding its receptor c-Met.NK4 is a four-kringle antagonist of HGF and exerts an antitumor activity,not only by HGF antagonism but also by antiangiogenesis.In order to clarify the antitumor activity of NK4 in ovarian cancer and investigate NK4 as a potential therapeutic method to ovarian cancer,NK4cDNA was induced into a human ovarian cancer cell line SKOV-3 and the effect of transfected NK4 gene on tumor growth of human ovarian cancer xenograft in nude mice.【Methods】 SKOV-3 cells were transfected with NK4 or luciferase expression plasmids.After antibiotics selected,NK4-expressing cells（SKOV-3/NK4） and control cells（SKOV-3/LUC） were obtained.Western blot analysis was used to detect NK4 expression in the supernatants of SKOV-3,SKOV-3/LUC and SKOV-3/NK4 cells.Western blot analysis was used to detect NK4 expression in the supernatants of SKOV-3,SKOV-3/LUC and SKOV-3/NK4 cells and to detect c-Met and phospho-c-Met expression in SKOV-3 cells cultured with supernatants from SKOV-3,SKOV-3/LUC and SKOV-3/NK4 cells.Cell growth curves were performed using MTT kit.Subcutaneous tumor growth curve were performed according to the xenograft tumor volume among the three groups.【Results】 NK4 was detected in the supernatant of SKOV-3/NK4 and NK4 expressed in the supernatant of SKOV-3/NK4 inhibited phosphorylation of c-Met in SKOV-3 cells.There was no inter-group difference among SKOV-3,SKOV-3/LUC or SKOV-3/NK4 cells in doubling time in vitro.The subcutaneous tumor volume in SKOV-3/NK4 group was significantly smaller than that in the control（SKOV-3 or SKOV-3/LUC） groups（P 〈0.05）.【Conclusion】 NK4 can be expressed by established SKOV-3/NK4 cells and NK4 can inhibit phosphorylation of c-Met in SKOV-3 cells.But NK4 has no effect on cell proliferation in SKOV-3 cells.NK4 can inhibit ovarian cancer growth in nude mice.These findings suggest NK4 can be a novel gene targeted therapeutic strategy to human ovarian cancer.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2012年第21期1-4,共4页
China Journal of Modern Medicine
基金
辽宁省博士启动基金项目(No:20111108)
关键词
卵巢癌
NK4
基因治疗
NK4； ovarian cancer； gene therapy；
