IPI-926对人结肠癌HT-29细胞增殖的影响及其机制  

Effect of IPI-926 on proliferation of human colon cancer HT-29 cells and its mechanism

作　　者：左赛[1] 毕研贞 孔令斌[3] 王一波[1] 张秋生[1] 洪丰[1] 张帆[1] 邱悦 ZUO Sai;BI Yan-zhen;KONG Ling-bin;et al(Clinical College of Jining Medical University, Jining, Shandong 272016, China)

机构地区：[1]济宁医学院附属医院消化内科,山东济宁272016 [2]首都医科大学附属北京佑安医院疑难肝病及人工肝中心,北京100000 [3]济宁医学院内科教研室,山东济宁272016 [4]济宁医学院,山东济宁272016

出　　处：《中华全科医学》2018年第2期173-175,322,共4页Chinese Journal of General Practice

基　　金：国家自然科学基金培育项目(JNYXYGZRPY2016-03);山东省济宁市科技局项目(JNSKJJ201555);济宁医学院大学生创新科研课题(JNYXYXSCX-201510)

摘　　要：目的观察IPI-926对人结肠癌HT-29细胞增殖的影响,并探讨在人结肠癌HT-29细胞增殖过程中IPI-926对Hedgehog(刺猬,Hh)信号通路因子表达的影响。方法常规体外培养人结肠癌HT-29细胞,随机分为对照组(空白培养基)和不同浓度IPI-926(10 nM、20 nM、30 nM)的实验组,培养HT-29细胞72 h后倒置显微镜观察各组细胞形态变化;MTT法测定各组培养72 h后HT-29细胞抑制率;RT-PCR和Western blot分别检测各组培养72 h后HT-29细胞Shh、Ptch、Smo和Gli-1 mRNA及其相关蛋白的表达;采用独立样本t检验进行组间比较,P<0.05为差异有统计学意义。结果对照组细胞生长状态良好;而用不同浓度IPI-926(10 nM、20 nM、30 nM)处理的HT-29细胞细胞凋亡增加,并随着IPI-926浓度的增加,凋亡率明显升高;10 nM、20 nM、30 nM浓度的IPI-926对人结肠癌细胞HT-29抑制率分别为(17.23±1.32)%、(38.34±1.82)%和(75.81±3.43)%,随着浓度的增加而升高,抑制率呈量效关系。空白对照组和IPI-926浓度为10 nM时Shh、Ptch、Smo和Gli-1的mRNA及其蛋白表达差异均无统计学意义,但在IPI-926浓度为20 nM和30 nM时,Shh、Ptch、Smo和Gli-1的mRNA及其蛋白表达均比空白对照组明显下调,并且IPI-926浓度为30 nM时的Shh、Ptch、Smo和Gli-1的表达明显高于20 nM时的mRNA表达。结论小剂量IPI-926不能有效抑制HT-29细胞的增殖,但当超过20 nM时则可以有效抑制HT-29细胞的增殖;其抑制的机制可能与Hh通路相关成员Shh、Ptch、Smo和Gli-1的表达有关。Objective To observe the effect of IPI-926 on the proliferation of human colon cancer HT-29 cells and to in- vestigate the effect of IPI-926 on the expression of Hedgehog signaling pathway in human colon cancer HT-29 cell prolifera- tion. Methods Human colon cancer HT-29 cells were cultured in vitro, randomly set the blank control group and different concentrations （10, 20, 30 nM） IPI-926 intervention group. morphological changes were observed by inverted microscope The intervention time of colorectal cancer cell was 72 h. The Effects of different concentrations of IPI-926 after the inter- vention of 72 h on the proliferation of HT-29 cells was determined by MTF assay and the apoptotic rate calculated. Reverse transcription PCR（RT-PCR） and Western blot were used to detect the expression of Shh, Ptch, Smo and Gli-1 mRNA and its related proteins in HT-29 cells after 72 hours of culture. The independent samples t-test was used for comparison be- tween groups, and P 〈 0.05 was considered statistically significant. Results Apoptosis of HT-29 cells treated with differ- ent concentrations of IPI-926 （10, 20, 30 nM） increased when compared with the control, and the apoptosis rate increased with the increase of IPI-926 concentration. The inhibitory rates of IPI-926 （ 10, 20 and 30 nM） on human colon cancer cell line HT-29 were （ 17.23 ±1.32） %, （ 38.34 ± 1.82） % and （75.81 ±3.43 ） %, respectively, rose with the increase of the concentration of IPI-926, showed a dose-effect relationship. The expression of Shh, Ptch, Smo and Gli-1 mRNA and pro- tein were not significantly different between the blank control group and the IPI-926 concentration of 10 nM. However, the mRNA and protein expression of Shh, Ptch, Smo and Gli-1 were significantly down-regnlated at IPI-926 concentration of 20 nM and 30 nM as compared with blank control group. And the expression of Shh, Ptch, Smo and Gli-1 at IPI-926 at 30 nM was significantly higher than that at 20 nM. Conclusion Low dose IPI-926 cannot effectively i

关 键 词：IPI-926 结肠癌 HT-29细胞 Hedgehog信号通路因子 

分 类 号：R735.35[医药卫生—肿瘤] R329.28[医药卫生—临床医学]