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目的研究溶血磷脂酸(lysophesphatidie acid,LPA)通过上调CXCL12-CXCR4的表达来促进卵巢癌的转移。方法培养卵巢癌细胞株CAOV3和SKOV3,免疫细胞化学方法检测两种细胞膜上的CXCR4蛋白表达。不同浓度的LPA刺激卵巢癌细胞,分别作用不同的时间后,采用逆转录-聚合酶链反应(RT-PCR)、流式细胞技术检测卵巢癌细胞膜上CXCR4的表达变化,采用ELISA方法检测细胞上清中CXCL12的变化。并建立卵巢癌裸鼠腹腔移植瘤模型,采用流式细胞检测及免疫组化技术测定瘤组织中的CXCR4的表达变化。结果卵巢癌细胞在细胞膜和细胞质中均有CXCR4蛋白表达,不同浓度和不同时间的LPA刺激细胞后,CXCR4和CXCL12的mRNA及蛋白表达水平均较无LPA组处理组明显升高,LPA处理组中移植瘤组织的CXCR4蛋白水平表达明显高于对照组,二者相比差异有显著性(P<0.01)。结论 LPA可以诱导卵巢癌细胞分泌CXCL12和CXCR4,因此LPA可以通过上调CXCL12-CXCR4生物轴的作用来促进卵巢癌的转移。
Objective To investigate the effect of lysophesphatidie acid (LPA) on the metastasis of ovarian cancer by up-regulating CXCL12-CXCR4 expression. Methods The ovarian cancer cell lines CAOV3 and SKOV3 were cultured and the expression of CXCR4 protein was detected by immunocytochemistry. Different concentrations of LPA stimulated ovarian cancer cells. After different time, the expression of CXCR4 in ovarian cancer cell membrane was detected by reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry, and the expression of CXCR4 was detected by ELISA Qing CXCL12 changes. The model of transplanted abdominal cavity in nude mice with ovarian cancer was established. The expression of CXCR4 in tumor tissue was detected by flow cytometry and immunohistochemistry. Results The expression of CXCR4 protein in both cell membrane and cytoplasm of ovarian cancer cells was increased. The mRNA and protein expressions of CXCR4 and CXCL12 in LPA-stimulated cells were significantly higher than those in non-LPA group The expression of CXCR4 protein in the xenografted tumor tissue was significantly higher than that in the control group, the difference was significant (P <0.01). Conclusion LPA can induce ovarian cancer cells to secrete CXCL12 and CXCR4. Therefore, LPA can promote ovarian cancer metastasis by up-regulating the biological axis of CXCL12-CXCR4.