二氢丹参酮调控Wnt/β-catenin信号通路抑制大肠癌细胞增殖的研究

Inhibitory effect of dihydrotanshinone on proliferation of colorectal cancer cells by regulating Wnt/β-catenin signaling pathways

目的研究二氢丹参酮对大肠癌细胞SW480增殖的抑制作用及对Wnt/β-catenin信号通路的调控。方法体外培养大肠癌细胞SW480,低、高剂量实验组分别予以5,15μmol·L^-1二氢丹参酮处理,对照组予以等量0. 9%Na Cl处理,分别干预24,48,72 h。以噻唑蓝(MTT)法、细胞克隆形成实验及5-溴脱氧尿嘧啶核苷(Brd U)实验检测SW480细胞增殖情况,以蛋白免疫印迹法检测Wnt/β-catenin信号通路相关蛋白表达情况。结果对照组及低、高剂量实验组SW480细胞的克隆形成率分别为(41. 65±5. 02)%,(26. 07±4. 23)%和(11. 49±2. 41)%,Brd U阳性细胞百分比分别为(64. 25±5. 78)%,(31. 07±3. 26)%和(19. 11±2. 01)%,β-catenin蛋白相对表达量分别为0. 95±0. 13,0. 97±0. 15和0. 92±0. 14,c-Myc蛋白相对表达量分别为1. 07±0. 18,0. 63±0. 12和0. 41±0. 09。低、高剂量实验组SW480细胞增殖抑制率随二氢丹参酮作用时间延长逐渐升高,且各干预时间点高剂量实验组均高于低剂量实验组(均P <0. 05);低、高剂量实验组SW480细胞克隆形成率、Brd U阳性细胞百分比及c-Myc蛋白相对表达量较对照组降低,且高剂量实验组低于低剂量实验组(均P <0. 05);3组β-catenin蛋白相对表达量差异无统计学意义(均P>0. 05)。结论二氢丹参酮可有效抑制大肠癌SW480细胞增殖,其作用机制与抑制c-Myc蛋白表达,阻碍Wnt/β-catenin信号转导相关。

Objective To explore the inhibition effect of dihydrotanshinone on the colorectal cancer cells SW480 proliferation and its regulation effect on Wnt/β-catenin signaling pathways. Methods Colorectal cancer cells SW480 were cultivated in vitro,experimental-L/H groups were treated with 5,15 μmol·L-1 dihydrotanshinone,control group was treated with equal amount of saline,and each group was treated for 24,48,72 h respectively. The proliferation of SW480 cells were detected by Methyl thiazolyl tetrazolium( MTT),cell clone forming experiments and5-bromine deoxidization uracil nucleoside( BrdU) experiment,the expression of Wnt/β-catenin signaling pathways related protein was detected by Western blot. Results The SW480 cell clone formation rates in control group and experimental-L/H groups were( 41. 65 ± 5. 02) %,( 26. 07 ± 4. 23) %,( 11. 49 ± 2. 41) %;the proportions of Brd U positive cell were( 64. 25 ± 5. 78) %,( 31. 07 ± 3. 26) %,( 19. 11 ± 2. 01) %;the β-catenin protein relative expression were 0. 95 ± 0. 13,0. 97 ± 0. 15,0. 92 ± 0. 14;the c-Myc protein relative expression were 1. 07 ± 0. 18,0. 63 ± 0. 12,0. 41 ± 0. 09. The SW480 cell proliferation inhibition rate increased gradually with the extended of dihydrotanshinone action time in experimental-L/H groups,and experimental-H group was higher than experimental-L group at each time point( P < 0. 05);the clone formation rate,proportion of BrdU positive cell and c-Myc protein relative expression decreased in experimental-L/H groups compared with control group,and experimental-H group was lower than experimental-L group( P < 0. 05);the β-catenin protein relative expression in 3 groups had no significant difference( P > 0. 05). Conclusion Dihydrotanshinone can inhibit the proliferation of colorectal cancer cells SW480,and the action mechanism may be related to the inhibition of c-Myc protein expression,and the blockade of signal transduction of Wnt/β-catenin.