摘要
目的在人卵巢癌细胞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/LUC和SKOV-3/NK4)体外生长速度,建立裸鼠卵巢癌腹水瘤模型,用Kaplan-Meier方法比较SKOV-3/LUC和SKOV-3/NK4细胞接种形成腹水瘤的裸鼠的累积生存率的差异。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表达在各组差异无统计学意义。三种细胞体外生长曲线比较差异无统计学意义(P>0.05)。腹腔内接种SKOV-3/LUC细胞的裸鼠,均产生腹腔内种植肿瘤伴腹水形成,于50 d内相继死亡,而接种SKOV-3/NK4细胞的裸鼠生存期均大于70 d,差异有统计学意义(P<0.01)。结论SKOV-3/NK4细胞在培养基中大量分泌NK4蛋白,NK4能抑制人卵巢癌细胞c-Met受体磷酸化。在裸鼠体内NK4通过抗血管生成作用抑制卵巢癌细胞腹腔内种植瘤的生长和腹水形成,提高累积生存率,改善预后。NK4可作为卵巢癌基因治疗的新方法。
Objective To learn the antitumor activity of NK4 in ovarian cancer and explore its potential therapeutic effect on ovarian cancer.Methods SKOV-3 cells were transfected with NK4 or luciferase expression plasmids to get the NK4-expressing cells（SKOV-3/NK4） and control cells（SKOV-3/LUC）.Western blot analysis was used to detect NK4 expression in the supernatants of SKOV-3,SKOV-3/LUC and SKOV-3/NK4 cells,as well as the c-Met and phospho-c-Met expression in SKOV-3 cells cultured with supernatants of SKOV-3,SKOV-3/LUC and SKOV-3/NK4 cells.Cell growth curves were obtained using MTT assay.Survival curves were obtained using Kaplan-Meier method after SKOV-3/LUC and SKOV-3/NK4 cells were injected intraperitoneally into nude mice.Results NK4 was detected in the supernatant of SKOV-3/NK4 and it inhibited the phosphorylation of c-Met in SKOV-3 cells.There was no intergroup difference in the expression of c-Met.All SKOV-3/LUC cell-inoculated mice died of peritoneal dissemination with ascites within 50 days after inoculation,in contrast all SKOV-3/NK4 cell-inoculated mice survived longer than 70 days（P0.01）.Conclusion NK4 can be expressed in the established SKOV-3/NK4 cells and inhibit the phosphorylation of c-Met in SKOV-3 cells.But NK4 has no effect on cell proliferation in SKOV-3 cells.NK4 can improve the accumulative survival rate of nude mice intraperitoneally inoculated ovarian cancer cell.These findings suggest that NK4 can be a novel gene targeted therapeutic strategy to human ovarian cancer predominantly by its anti-angiogenesis function via inhibiting HGF-c-Met signaling pathway or other putative pathway.
出处
《中国微生态学杂志》
CAS
CSCD
2012年第12期1071-1073,共3页
Chinese Journal of Microecology
基金
辽宁省博士启动课题(20111108)
关键词
卵巢癌
NK4
基因治疗
累计生存率
Ovarian cancer
NK4
Gene therapy
Accumulative survival rate
