本文選題：膀胱腫瘤 + Wnt信號通路�。� 參考：《腫瘤》2017年11期

【摘要】：目的 :分析dickkopf 2(DKK2)基因在人膀胱癌細(xì)胞系及膀胱癌組織中的表達(dá)水平,以及DKK2過表達(dá)對人膀胱癌細(xì)胞增殖與遷移的影響,并初步探討其作用機(jī)制。方法:通過RT-PCR、實(shí)時(shí)熒光定量PCR和蛋白質(zhì)印跡法檢測人膀胱癌細(xì)胞系及組織中DKK 2基因的相對表達(dá)水平。選用DKK2低表達(dá)的膀胱癌細(xì)胞系T24,進(jìn)行DKK2過表達(dá)質(zhì)粒轉(zhuǎn)染后,采用RT-PCR、實(shí)時(shí)熒光定量PCR和蛋白質(zhì)印跡法驗(yàn)證DKK2過表達(dá)效果;然后采用CCK-8法、平板克隆形成實(shí)驗(yàn)、劃痕愈合實(shí)驗(yàn)、Transwell小室法和FCM法分別檢測DKK2過表達(dá)對細(xì)胞增殖、克隆形成、遷移、侵襲及細(xì)胞周期的影響;進(jìn)一步采用蛋白質(zhì)印跡法和實(shí)時(shí)熒光定量PCR法檢測DKK2過表達(dá)后膀胱癌細(xì)胞中增殖和遷移相關(guān)分子表達(dá)水平的變化。結(jié)果:人膀胱癌細(xì)胞系5637、T24、SW780、J82、HT-1197和HT-1376中DKK2 mRNA及蛋白表達(dá)水平均明顯低于正常膀胱上皮細(xì)胞SVHUC-1(P值均0.01);同時(shí)與正常膀胱組織及配對的癌旁組織相比,膀胱癌組織中DKK2 mRNA及蛋白的表達(dá)均明顯下調(diào)(P值均0.001)。DKK2過表達(dá)質(zhì)粒轉(zhuǎn)染后,膀胱癌T24細(xì)胞中DKK2 mRNA及蛋白的表達(dá)水平明顯上調(diào),而細(xì)胞的增殖、克隆形成、遷移和侵襲能力均受到明顯抑制(P值均0.05),同時(shí)細(xì)胞周期被阻滯于G0/G1期(P0.001)。而且DKK2過表達(dá)的膀胱癌T24細(xì)胞中,p21和E-cadherin表達(dá)水平明顯升高(P值均0.001),cyclin D1、vimentin和N-cadherin表達(dá)水平明顯降低(P值均0.001)。結(jié)論:DKK2在人膀胱癌細(xì)胞及組織中低表達(dá)。DKK 2可能作為一個(gè)潛在的抑癌基因,參與膀胱癌進(jìn)展。
[Abstract]:Aim: to investigate the expression of dickkopf 2 gene in human bladder cancer cell lines and bladder cancer tissues, and the effect of DKK2 overexpression on the proliferation and migration of human bladder cancer cells. Methods: RT PCR, real-time quantitative PCR and Western blot were used to detect the relative expression of DKK 2 gene in human bladder cancer cell lines and tissues. Bladder cancer cell line T24, which has low expression of DKK2, was used to transfect DKK2 overexpression plasmid, RT-PCR, real-time fluorescence quantitative PCR and Western blotting were used to verify the effect of DKK2 overexpression. The effects of DKK2 overexpression on cell proliferation, clone formation, migration, invasion and cell cycle were detected by FCM and Transwell chamber method. Western blotting and real-time fluorescence quantitative PCR were used to detect the expression of proliferative and migration-related molecules in bladder cancer cells after overexpression of DKK2. Results: the expression levels of DKK2 mRNA and protein in human bladder cancer cell line 5637, T24T24, SW780, J82, HT-1197 and HT-1376 were significantly lower than those in normal bladder epithelial cells, and were significantly lower than those in normal bladder tissues and matched adjacent tissues. The expression of DKK2 mRNA and protein in bladder cancer tissue was significantly down-regulated, and the expression of DKK2 mRNA and protein in T24 cells was upregulated, while the proliferation and clone formation of T24 cells were observed after transfection with both 0.001).DKK2 overexpression plasmids. Migration and invasion were significantly inhibited (P = 0.05) and cell cycle was blocked in G0/G1 phase (P 0.001). Furthermore, the expression of p21 and E-cadherin in T24 cells with overexpression of DKK2 was significantly increased (P = 0.001), and the expression of cyclin D _ (1) vimentin and N-cadherin decreased significantly (P = 0.001). Conclusion the low expression of DKK2 in human bladder cancer cells and tissues may be a potential tumor suppressor gene involved in the progression of bladder cancer.
【作者單位】： 重慶市中醫(yī)院(重慶第一人民醫(yī)院)腎內(nèi)科;
【分類號】：R737.14

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