Shh信号通路阻断对人结肠癌细胞HT-29增殖、侵袭的影响及机制探讨

Effects of blocking Shh signaling pathway on proliferation and invasion of colon cancer cell line HT-29

目的观察阻断Sonic Hedgehog(Shh)信号通路对人结肠癌细胞HT-29增殖和侵袭的影响,并探讨其机制。方法取对数生长期HT-29细胞分为A、B、C、D组,分别置于含0、5、10、20μmol/L Shh信号通路特异性抑制剂Cyclopamine的培养基中培养。采用MTT法检测各组干预24、48、72 h时细胞增殖抑制率,采用Transwell法检测各组干预48 h时体外侵袭能力,采用实时荧光定量PCR法检测各组细胞Shh、下游转录因子GLI1 mRNA。结果干预24、48、72 h时各组细胞增殖抑制率为B组<C组<D组,P均<0.05;A、B、C、D组的穿膜细胞数分别为(55.670±6.506)、(35.330±7.371)、(28.330±4.041)、(23.330±2.309)个,B、C、D组穿膜细胞数均低于A组,P均<0.05,但B、C、D组间比较差异无统计学意义。HT-29细胞Shh、GLI1 mRNA的相对表达量均为A组>B组>C组>D组,P均<0.05。结论阻断Shh信号通路可抑制HT-29细胞增殖、降低体外侵袭能力,可能与其降低Shh、GLI1表达有关。

Objective To obseirve the effects of blocking Sonic Hedgehog （Shh） signaling pathway on the proliferation and invasion of colon cancer cell line HT-29 and to investigate the mechanism. Methods HT-29 cells in the logarithmic phase were divided into groups A, B, C, and D and were cultured in the culture medium containing 0, 5, 10 and 20 mol/ L Shh signaling pathway specific inhibitor Cyclopamine, respectively. The inhibitory rate of cell proliferation was detected by MTT method at 24, 48 and 72 h after the treatment of Cyclopamine. Transwell method was used to detract the invasive a- bility of each group at 48 h. The Shh and downstream transcription factor GLI1 mRNA in HT-29 ceils was detected by real- time fluorescence quantitative PCR. Results The proliferation inhibition rate at 24 h, 48 h and 72 h ： group B 〈 group C 〈 group D, all P 〈 0.05. The number of transmembrane Ceils in groups A, B, C and D was 55. 670 ± 6.506, 35. 330 ± 7. 371, 28. 330 ±4. 041 and 23. 330 ±2. 309, respectively. The number of transmembrane cells in the groups B, C and D was lower than that of group A, all P 〈 0.05, but there was no significance difference between the groups B, C and D. The relative expression of Shh and GLI1 mRNA in HT-29 cells was group A 〉 group B 〉 group C 〉 group D, all P 〈0.05. Con- clusion Blocking Shh signaling pathway can inhibit the proliferation of HT-29 ceils and decrease the invasion ability in vitro, which may be related to the decreased expression of Shh and GLI1.