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目的研究中药消癌平注射液(xiaoaiping injection,XAP)对卵巢癌细胞Caov-3侵袭的作用及其可能机制。方法向体外培养的卵巢癌细胞株Caov-3中加入胎牛血清(FBS),同时加入消癌平注射液,①利用肿瘤细胞侵袭实验观察消癌平注射液是否具有抑制卵巢癌细胞侵袭性增加的作用,②采用荧光染色法观察微丝(f-actin)的重排,③采用蛋白质免疫印迹实验检测影响微丝重排的信号分子的表达。并且,在整个实验过程中应用磷脂酰肌醇3-激酶(PI3K)特异性抑制剂LY294002作为阳性对照。结果 Caov-3细胞在FBS诱导下,穿过Transwell的细胞数明显增加;荧光显微镜下可观察到细胞微丝f-actin发生重新排布,形成了片状伪足;PI3K激酶的下游分子丝氨酸/苏氨酸蛋白激酶Akt磷酸化表达增加。在FBS诱导的同时加入LY294002,则可抑制Caov-3细胞发生上述改变;同样,在FBS诱导的同时加入消癌平注射液,其干预作用与LY294002相似,也可降低Caov-3细胞的侵袭性、抑制微丝actin的重排及片状伪足的形成、抑制的Akt磷酸化。结论消癌平注射液具有抑制PI3K激酶活性,抑制卵巢癌Caov-3细胞微丝actin重排、抑制细胞侵袭的作用。
Objective To investigate the effect of xiaoaiping injection (XAP) on the invasion of ovarian cancer cells Caov-3 and its possible mechanism. Methods FBS was added to ovarian cancer cell line Caov-3 cultured in vitro. At the same time, Xiaofanping injection was added into the ovarian cancer cell line in vitro. ① The tumor invasion experiment was used to observe whether Xiaojianping injection could inhibit the invasiveness of ovarian cancer cells (2) Fluorescent staining was used to observe the rearrangement of f-actin, (3) Western blotting was used to detect the expression of signaling molecules that influence the microwire rearrangement. Also, phosphatidylinositol 3-kinase (PI3K) -specific inhibitor LY294002 was used as a positive control throughout the experiment. Results The cell number of Caov-3 cells translocated through Transwell was significantly increased under the induction of FBS. Fluorescence microscopy showed that f-actin was rearranged to form lamellipodia. The downstream molecular serine / Threonine protein kinase Akt phosphorylation increased. Similarly, the addition of LY294002 to FBS-induced Caov-3 cells inhibited the above changes; similarly, the addition of Xiaofanping injections induced by FBS had similar effects as LY294002 in reducing the invasiveness of Caov-3 cells , Inhibit the actin filaments rearrangement and the formation of lamellipodia, inhibition of Akt phosphorylation. Conclusion Xiaojianping injection can inhibit PI3K kinase activity and inhibit actin rearrangement and inhibit cell invasion in ovarian cancer Caov-3 cells.