MSP影响非小细胞肺癌PC14细胞周期和上皮间质转化的研究

Effects of MSP on Cell Cycle and EMT of Non-Small Cell Lung Cancer PC14

目的研究巨噬细胞刺激蛋白(MSP)在无巨噬细胞刺激蛋白受体(RON)的情况下对非小细胞肺癌PC14细胞周期的影响,并分析其对PC14上皮间质转化(EMT)的影响。方法取MSP阴性、RON阴性的PC14细胞系,稳定表达MSP的细胞系PC14-Mst1-pEGFP-N1和稳定表达荧光蛋白的空载质粒细胞系PC14-pEGFPN1,流式细胞术检测各细胞系细胞周期的影响;透射电镜观察在细胞增殖期间细胞之间的连接间隙;逆转录(RT)-PCR和Western blot分别检测E-钙粘蛋白(E-cadherin)和波形蛋白(Vimentin)mRNA、蛋白表达水平。结果流式细胞术结果显示与PC14、PC14-pEGFP-N1相比,PC14-Mst1-pEGFP-N1细胞G_1/G_0期数量增多,S期与G_2/M期减少;透射电镜发现PC14-Mst1-pEGFP-N1细胞之间的细胞连接较紧密,细胞之间空隙距离缩小;RT-PCR和Western blot检测结果显示,与PC14细胞相比,PC14-Mst1-pEGFP-N1细胞E-cadherin的mRNA和蛋白表达水平升高,而Vimentin的mRNA、蛋白表达水平降低。结论 MSP在RON阳性的情况下,可使G_1期肺癌细胞增多、分裂相减少,抑制EMT。

Objective To study the effect of macrophage stimulating protein （MSP） on the cell cycle of non- small cell lung cancer PC14 cells without expression of recepteur d＇originenanta （RON） and MSP, and analyse its effect on PC14＇s epithelial mesenehymal transition （EMT） capacity. Methods Vitro culture PC14 （blank control）, PC14-Mstl-pEGFP-N1 （stablely expressed MSP） and PC14-pEGFP-N1. Cell cycles were detected by flow cytometry and the gaps between cells during growth were measured by transmission electron microscope （TEM） ; RT-PCR and Western blot were used to figure out the shifts of EMT related gene expression in PC14-Mstl-pEGFP- N1 ceils. Results Compared with the PC14 group and PC14-pEGFP-N1 group, PC14-Mstl-pEGFP-N1 population of G1/G0 phase were significantly increased while S and G2/M phase were significantly reduced; The gaps between PC14-Mstl-pEGFP-N1 cells decreased; RT-PCR and Western blot showed that mRNA and protein levels of E- cadherin of PC14-Mstl-pEGFP-N1 were significantly higher than that of PC14, but mRNA and protein levels of Vimentin were significantly lower. Conclusion MSP may affect the cell cycle of PC14 and inhibit its EMT procedure by regulating the expression of related proteins including E-cadherin and Vimentin when RON was not expressed.