Taqman qPCR检测CD147/Basigin剪接变异体在人上皮性卵巢癌中的表达

Quantification of CD147/basigin Splicing Variants by Taqman qPCR in Epithelium Ovarian Cancer

目的:应用Taqman qPCR技术检测CD147/basigin剪接变异体在人上皮性卵巢癌组织与正常卵巢组织中的表达差异。方法:运用半定量RT-PCR技术检测CD147/basigin剪接变异体在上皮性卵巢癌细胞系中的表达;Taqman qPCR检测CD147/basigin剪接变异体在人上皮性卵巢癌细胞系中的表达分布;进一步通过收集32例上皮性卵巢癌组织与26例正常卵巢组织,提取组织RNA,反转录cDNA,Taqman qPCR检测CD147/basigin剪接变异体mRNA在上皮性卵巢癌组织与正常卵巢组织中的表达差异。结果:半定量RT-PCR结果显示basigin-2,basigin-3和basigin-4在上皮性卵巢癌细胞系中均有表达,主要以basigin-2为主;Taqman qPCR检测到三种剪接变异体在不同卵巢癌细胞系中表达不同,basigin-2在卵巢癌细胞系中较basigin-3,basigin-4表达较高,basigin-4较basigin-3略高;Basigin-2剪接变异体在高转移Ho-8910pm细胞中表达较高,在低转移HO-8910细胞中表达较低。组织Taqman qPCR检测basigin-2和basigin-4在上皮性卵巢癌组织中的表达水平显著高于正常卵巢组织(P值分别为<0.0001和0.0261),basigin-3的表达水平略有升高(P=0.2616),但无统计学意义。结论:三种剪接变异体在卵巢癌组织中较正常卵巢组织表达上调。CD147/basigin-2在高转移卵巢癌细胞系HO-8910pm中高表达,在低转移卵巢癌细胞系HO-8910中低表达,且表达强度与上皮性卵巢癌的转移相关;探讨CD147/basigin-2在上皮性卵巢癌中的高表达,为卵巢癌的进一步治疗开辟一新途径。

Objective： To determine the expression differences of CD147/basigin splice variants between epithelial ovarian cancer tissue and normal ovarian tissue by Taqman qPCR. Methods： Semi-quantitative qPCR was used to detect splice variants of CD147/basi- gin expression level, and Taqman qPCR was used to analyze their distribution in epithelial ovarian cancer cell lines. Furthermore, RNA was extracted from 32 cases of epithelial ovarian cancer and 26 cases of normal ovarian tissue, reversed into eDNA, and then the expres- sion difference of CD147/basigin splicing variant mRNA between epithelial ovarian cancer tissues and normal ovarian tissue was deter- mined by Taqman qPCR. Results： Semi-quantitative qPCR results show that all of Basigin-2, Basigin-3 and Basigin-4 express in epithe- lial ovarian cancer cell lines, but mainly Basigin-2. Taqman qPCR results demonstrate that three splice variants express differently in dif- ferent ovarian cancer cell lines： the expression level of Basigin-2 is the highest, and Basigin-4 is higher than basigin-3 in ovarian cancer cell lines; Basigin-2 expression level is higher in highly metastatic Ho-8910pm cells, and lower in lowly metastatic HO-8910 cells. Tiss- sue Taqman qPCR shows that basigin-2 and basigin-4 expression in epithelial ovarian cancer tissue are significantly higher than in the normal ovarian tissue （P values were 〈0.0001 and 0.0261）, basigin-3 is slightly higher （P = 0.2616）, which is of no statistically signifi- cance. Conclusion： The three splice variants are up-regulated in ovarian cancer tissue compared with normal ones. And CD147/basigin-2 expression level is related to the metastasis of epithelial ovarian cancer. The exploration for mechanism of high expression level of CD 147/basigin-2 in epithelial ovarian cancer paves a new path for ovarian cancer treatment.