淋巴管内皮细胞微环境对卵巢癌淋巴结定向高转移细胞上皮-间质转化的影响

Ability of lymphatic endothelial cell microenvironment to affect the epithelial-mesenchymal transition in ovarian carcinoma cell lines showing highly lymphatic directional metastasis

目的通过人卵巢癌淋巴结定向高转移的细胞株(SKOV3-PM4)与人淋巴管内皮细胞株(HLEC)的条件培养和共培养,探讨HLEC微环境对SKOV3-PM4的运动迁移能力以及上皮-间质转化(EMT)的影响。方法采用transwell小室体外侵袭法,观察经HLEC的条件培养后SKOV3-PM4的侵袭、迁移能力的改变。SKOV3-PM4经HLEC的条件培养和共培养后的爬片细胞,采用免疫组化法检测转化生长因子(TGF-β1)、E-钙黏蛋白(E-cadherin)、波形蛋白(vimentin)表达的变化。结果经HLEC培养基的条件培养后SKOV3-PM4细胞的侵袭率为(53±11)%,单独培养的SKOV3-PM4细胞的侵袭率为(30±5)%(P<0.05);条件培养后SKOV3-PM4细胞的迁移率为(48±5)%,单独培养的SKOV3-PM4细胞的迁移率为(34±2)%(P<0.05)。SKOV3-PM4细胞经条件培养和共培养后TGF-β1和vimentin表达上调(P均<0.05),E-cadherin表达下调(P<0.05)。结论淋巴管内皮细胞微环境可促进卵巢癌淋巴结定向高转移细胞间质化的转变,增强卵巢癌细胞的侵袭、迁移能力。

Objective To explore whether creating a human lymphatic endothelial cell microenvironment under culture or co-culture conditions can affect the invasion and migration activity and the epithelial-mesenchymal transition in the ovarian carcinoma SKOV3- PM4 cell line,which shows highly lymphatic directional metastasis. Methods SKOV3-PM4 cells were cultured in a lymphatic endothelial cell microenvironment either by using conditioned medium from cultures of human lens epithelium cells（HLECs） or by co- culturing with HLECs.The ability of SKOV3-PM4 ceils to invade and migrate was observed using transwell chambers.Expression of transforming growth factor （TGF-~t）,E-cadherin and vimentin in SKOV3-PM4 cells was analyzed by immunohistochemistry. Results The proportion of SKOV3-PM4 cultures showing invasion after incubation in HLEC-conditioned medium was（53＋l 1 ）% ,compared to （30＋5）% for control cells;the proportion of SKOV3-PM4 cultures showing migration behavior was（48＋5）%,compared to（34＋2）% for control cells.Expression of TGF-~I and vimentin was up-regulated and expression of E-cadherin down-regulated in SKOV3-PM4 cells after culturing them in HLEC-conditioned medium or co-culturing them with HLECs. Conclusions A human lymphatic endothelial cell microenvironment can alter the epithelial-mesenchvmal transition in ovarian carcinoma cells and enhance their invasion and migration ability.