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目的观察溶血磷脂酸(Lysophosphatidic acid,LPA)对人卵巢癌细胞SKOV3 P120catenin(P120ctn)表达的影响,探讨LPA和P120ctn在卵巢癌发生发展中的作用机制。方法将不同浓度的LPA作用于SKOV3细胞,MTT法检测细胞的增殖率;以最佳浓度的LPA及不同浓度的酪氨酸蛋白激酶抑制剂Genistein作用于SKOV3细胞,MTT法检测细胞的增殖抑制率;将SKOV3细胞分为3组:对照组、LPA组和LPA+Genistein组,分别于培养6、12、24h,采用免疫荧光细胞化学法和Western blot分析P120ctn表达的变化,RT-PCR法检测P120ctn基因mRNA的转录水平。结果 20μmol/LLPA作用24h促进SKOV3细胞增殖的作用最强,而200μmol/LGenistein对LPA促进SKOV3细胞增殖有明显的抑制作用。LPA可诱导SKOV3细胞膜中P120ctn的表达减少,胞质中的表达增多;而Genistein与LPA共同作用后,P120ctn在细胞膜中的表达增多,而在胞质中的表达减少。结论 LPA可诱导SKOV3细胞中P120ctn表达的改变,其机制可能与相关的酪氨酸蛋白激酶磷酸化有关。
Objective To investigate the effect of Lysophosphatidic acid (LPA) on the expression of human ovarian cancer cell line SKOV3 P120ctn and explore the mechanism of LPA and P120ctn in the development and progression of ovarian cancer. Methods Different concentrations of LPA were applied to SKOV3 cells and MTT assay was used to detect the proliferation of SKOV3 cells. The best concentration of LPA and different concentration of tyrosine kinase inhibitor Genistein were applied to SKOV3 cells. MTT assay was used to detect the proliferation inhibition rate ; The SKOV3 cells were divided into 3 groups: control group, LPA group and LPA + Genistein group, cultured for 6, 12 and 24 hours respectively. The changes of P120ctn expression were analyzed by immunofluorescence cytochemistry and Western blot. The expression of P120ctn Gene mRNA transcription level. Results The effect of 20μmol / LLPA for 24 hours was the strongest on SKOV3 cells, while 200μmol / LGenistein significantly inhibited the proliferation of SKOV3 cells induced by LPA. LPA could induce a decrease of P120ctn expression in the membrane of SKOV3 cells and an increase in the expression of the cytoplasm. However, with the combination of Genistein and LPA, the expression of P120ctn increased in the cell membrane and decreased in the cytoplasm. Conclusion LPA can induce the change of P120ctn expression in SKOV3 cells, which may be related to the phosphorylation of tyrosine kinase.