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目的探讨缺氧在卵巢上皮性癌细胞系SKOV3和ES2细胞拟态血管形成过程中的作用及西罗莫司对其的抑制效应。方法建立人工基底膜基质凝胶Matrigel三维培养系统,分别在常氧(常氧组)、缺氧(缺氧组)、缺氧时加入西罗莫司(缺氧+西罗莫司组)等条件下诱导,观察SKOV3和ES2细胞形成拟态血管的能力;应用相差显微镜和扫描电镜观察拟态血管的形态及其立体结构的特点;用RT-PCR技术检测不同条件下缺氧诱导因子(HIF)1αmRNA的相对表达量。结果培养7d后,缺氧组SKOV3和ES2细胞出现变形、伸展,形成腔样、管状、分支和网络状等管道结构;而常氧组及缺氧+西罗莫司组,部分细胞出现伸展但不形成管道结构。缺氧组SKOV3和ES2细胞的HIF-1αmRNA表达量分别为0·801±0·034和0·736±0·059,显著高于缺氧+西罗莫司组(分别为0·025±0·007和0·231±0·035;P<0·01,P<0·05)和常氧组(分别为0·010±0·004和0·011±0·002;P均<0·01)。结论缺氧是卵巢上皮性癌细胞形成拟态血管的重要诱导因素,西罗莫司通过抑制HIF-1αmRNA的表达阻断拟态血管的形成。
Objective To investigate the role of hypoxia in mimicry angiogenesis in epithelial ovarian cancer cell lines SKOV3 and ES2 and its inhibitory effect on sirolimus. Methods Matrigel three-dimensional culture system of artificial basement membrane matrix gel was established. The cells were exposed to sirolimus (hypoxia + sirolimus group) in normoxia group, hypoxia group and hypoxia group respectively Under the conditions of SKOV3 and ES2 cells were observed to form mimicry vascular ability; observed by phase contrast microscopy and scanning electron microscopy mimicry vascular morphology and its three-dimensional structure of the characteristics; using RT-PCR technology to detect hypoxia inducible factor (HIF) 1α mRNA Relative expression level. Results After cultured for 7 days, the SKOV3 and ES2 cells in the hypoxia group deformed and stretched and formed the tube-like structures of cavities, tubes, branches and networks. In the normoxia group and the hypoxia plus sirolimus group, some cells appeared to stretch Does not form a pipe structure. The expression of HIF-1αmRNA in SKOV3 and ES2 cells in hypoxia group were 0 · 801 ± 0 · 034 and 0 · 736 ± 0 · 059, respectively, which were significantly higher than those in hypoxia + sirolimus group (0 · 025 ± 0 · 007 and 0 · 231 ± 0 · 035; P <0.01, P <0.05) and normoxia groups (0 · 010 ± 0 · 004 and 0 · 011 ± 0 · 002, respectively; all P <0 · 01). Conclusions Hypoxia is an important inducing factor for the formation of mimic vessels in epithelial ovarian cancer cells. Sirolimus blocks the formation of mimic vessels by inhibiting the expression of HIF-1α mRNA.