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目的观察聚腺苷酸二磷酸核糖聚合酶-1(PARP-1)在肝细胞生长因子(HGF)诱导的卵巢癌细胞侵袭转移中的作用及相关信号转导通路。方法 Transwell实验检测不同浓度HGF对卵巢癌SKOV-3细胞体外侵袭力的影响;实时荧光定量PCR、Western blotting检测不同浓度及作用时间HGF对SKOV-3细胞PARP-1m RNA及蛋白表达的影响;RNA干扰沉默SKOV-3细胞的PARP-1基因,按不同处理因素将SKOV-3细胞分为空白对照组、HGF组、PARP-si RNA组、HGF+PARP-si RNA组;NC-si RNA组、HGF+NC-si RNA组。Western blotting检测不同处理组细胞内PARP-1蛋白表达。Transwell实验检测各组细胞侵袭力。ELISA法检测各组细胞上清中基质金属蛋白酶-2(MMP-2)的含量。结果 HGF可明显促进SKOV-3细胞的体外侵袭能力,并呈浓度时间依赖性。40 ng/m L HGF作用24 h后,HGF可明显上调SKOV-3细胞中PARP-1的表达水平;PARP-si RNA组细胞PARP-1的表达低于NC-si RNA组(P<0.05),干扰PARP-1的表达降低了HGF对SKOV-3细胞侵袭力及M M P-2含量的影响。结论HGF可促进卵巢癌SKOV-3细胞的侵袭转移,其作用机制可能与PARP-1介导的M M P-2表达增加有关。
Objective To investigate the role of PARP-1 in invasion and metastasis of ovarian cancer cells induced by hepatocyte growth factor (HGF) and related signal transduction pathway. Methods Transwell assay was used to detect the effect of different concentrations of HGF on invasiveness of ovarian cancer SKOV-3 cells in vitro. Real-time quantitative PCR and Western blotting were used to detect the effect of different concentrations of HGF on PARP-1mRNA and protein expression in SKOV-3 cells. The SKOV-3 cells were divided into blank control group, HGF group, PARP-si RNA group, HGF + PARP-si RNA group and NC-si RNA group according to different treatment factors, HGF + NC-si RNA group. Western blotting was used to detect the expression of PARP-1 in different treatment groups. Transwell assay detected cell invasiveness in each group. ELISA method was used to detect the content of matrix metalloproteinase-2 (MMP-2) in the supernatant of each group. Results HGF significantly promoted the invasion of SKOV-3 cells in vitro and in a concentration-dependent manner. HGF up-regulated the expression of PARP-1 in SKOV-3 cells after treated with 40 ng / ml HGF for 24 h. The PARP-1 expression in PARP-si RNA group was lower than that in NC-si RNA group (P <0.05) , Interfering with the expression of PARP-1 decreased the invasiveness of SKOV-3 cells and the effect of HGF on the content of MM P-2. Conclusions HGF can promote the invasion and metastasis of ovarian cancer SKOV-3 cells, which may be related to the increase of M P-2 expression induced by PARP-1.