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3维体外培养法(Three dimensional culture)在研究某些疾病发生发展的中存在重要意义.本研究运用3维悬滴培养技术,观察前列腺良性上皮细胞系RWPE-1和BPH-1,前列腺癌细胞系PC3、LNCaP、LNCaPable和Du145在3D悬滴培养条件下的形态学特征.运用实时定量RT-PCR技术比较了在普通2D培养与3D悬滴培养条件下E-钙黏连蛋白(E-cadherin)、波形蛋白(Vimentin)、雌激素受体α与β(ERα,ERβ),G蛋白偶联受体(GPR30)以及雄激素受体(AR)的表达变化.结果表明,与2D培养法相比,3D悬滴培养法抑制了前列腺上皮细胞发生EMT;在RWPE-1、BPH-1和PC-3细胞系中上调了ERα的表达,在LNCaP与LNCaP-able中下调了AR的表达.提示,与2D法相比3D悬滴培养法培养的前列腺上皮细胞具有不同的生物学特性.
Three-dimensional culture is of great significance in the study of the development of certain diseases.In this study, 3-dimensional hanging drop culture was used to observe the expression of RWPE-1 and BPH-1 in prostate benign epithelial cells, The morphological characteristics of PC3, LNCaP, LNCaPable and Du145 in 3D hanging drop culture were studied by real-time quantitative RT-PCR. The expression of E-cadherin ), Vimentin, ERα, ERβ, GPR30 and androgen receptor (AR) were detected by immunohistochemistry.The results showed that compared with 2D culture , 3D hanging drop culture inhibited the EMT of prostate epithelial cells, upregulated the expression of ERα in RWPE-1, BPH-1 and PC-3 cell lines, and downregulated the expression of AR in LNCaP and LNCaP-able, Prostate epithelial cells cultured by 3D hanging drop culture method have different biological characteristics compared with 2D method.