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目的 :探讨姜黄素对转化生长因子β1(transforming growth factor-beta 1,TGF-β1)诱导的食管癌KYSE70细胞上皮-间质转化(epithelialmesenchymal transformation,EMT)的影响。方法:不同浓度姜黄素处理KYSE70细胞12、24、48和72 h后,应用CCK-8法检测细胞的增殖情况。应用姜黄素(20μmol/L)、TGF-β1(10ng/m L)和姜黄素(20μmol/L)联合TGF-β1(10 ng/m L)处理KYSE70细胞后,在光学显微镜下观察细胞的形态,Transwell小室法检测各组细胞的侵袭能力,FCM法检测各组细胞的细胞周期分布,实时荧光定量PCR法和蛋白质印迹法分别检测各组细胞中EMT相关标志物E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)和波形蛋白(vimentin)m RNA及蛋白的表达。结果 :不同浓度姜黄素作用不同时间均能抑制食管癌KYSE70细胞的增殖(P值均<0.01)。TGF-β1处理后,食管癌KYSE70细胞形态向间质细胞转变,呈长梭形,但这种转变可以被姜黄素抑制。TGF-β1处理组细胞中G0/G1期细胞所占百分比低于对照组(KYSE70细胞未进行任何处理)(P=0.021);姜黄素联合TGF-β1处理组细胞中S期细胞所占百分比高于TGF-β1处理组(P<0.001),而G_2/M期细胞所占百分比低于TGF-β1处理组(P<0.001)。TGF-β1处理组的细胞侵袭数多于对照组(P<0.001),姜黄素处理组的细胞侵袭数少于对照组(P<0.001),姜黄素联合TGF-β1处理组的细胞侵袭数少于TGF-β1处理组(P<0.001)。TGF-β1处理组细胞中E-cadherin m RNA(P=0.045)和蛋白(P=0.008)的表达水平均低于对照组,而N-cadherin和vimentin m RNA(P=0.003和P<0.001)及蛋白(P值均<0.001)的表达水平均高于对照组;姜黄素联合TGF-β1处理组细胞中E-cadherin m RNA(P=0.005)和蛋白(P=0.006)的表达水平均高于TGF-β1处理组,而N-cadherin和vimentin m RNA(P=0.010和P<0.001)及蛋白(P值均<0.001)的表达水平均低于TGF-β1处理组。结论 :姜黄素可抑制食管癌KYSE70细胞的增殖和侵袭,还可有效抑制EMT的发生。
Objective: To investigate the effect of curcumin on the epithelialmesenchymal transformation (EMT) induced by transforming growth factor-β1 (TGF-β1) in esophageal carcinoma KYSE70 cells. Methods: KYSE70 cells were treated with different concentrations of curcumin for 12,24,48 and 72 h. CCK-8 assay was used to detect the proliferation of KYSE70 cells. KYSE70 cells were treated with curcumin (20μmol / L), TGF-β1 (10ng / m L), curcumin (20μmol / L) and TGF-β1 (10ng / Transwell chamber assay was used to detect the invasion ability of each group. The cell cycle distribution of each group was detected by FCM. The expression of EMT-related marker E-cadherin (E-cadherin) in each group was detected by real-time fluorescence quantitative PCR and Western blotting. cadherin, N-cadherin and vimentin m RNA and protein expression. Results: Different concentrations of curcumin inhibited the proliferation of KYSE70 cells at different times (all P <0.01). TGF-β1 treatment, esophageal cancer KYSE70 cell morphology changes to stromal cells, spindle fusiform, but this change can be curcumin inhibition. The percentage of G0 / G1 phase cells in TGF-β1 treated group was lower than that in control group (KYSE70 cells were not treated) (P = 0.021). The percentage of S phase cells in curcumin combined with TGF-β1 treatment group was high (P <0.001), while the percentage of G 2 / M phase cells was lower than that of TGF-β 1 -treated group (P <0.001). The number of cell invasion in the TGF-β1 treated group was more than that in the control group (P <0.001). The number of cell invasion in the curcumin-treated group was less than that in the control group (P <0.001) TGF-β1-treated group (P <0.001). The expression levels of E-cadherin m RNA (P = 0.045) and protein (P = 0.008) in TGF-β1 treated group were lower than those in control group, while N-cadherin and vimentin m RNA (P = 0.006) were higher than that of the control group (P <0.001). The expression of E-cadherin m RNA and protein in the cells treated with curcumin and TGF- TGF-β1 treatment group, while the expression levels of N-cadherin and vimentin m RNA (P = 0.010 and P <0.001) and protein (P <0.001) were lower than that of TGF-β1 treatment group. CONCLUSION: Curcumin can inhibit the proliferation and invasion of esophageal cancer KYSE70 cells and inhibit the occurrence of EMT effectively.