雷替曲塞及其联合PD98059对HT-29细胞的影响

目的 研究雷替曲塞及其联合PD98059对结肠癌细胞HT-29增殖及凋亡的影响,并探讨其分子机制.方法 ① 在倒置显微镜下观察处理24 h后各组细胞的形态及数量变化;② 使用细胞计数法检测不同处理条件下的细胞数;③ 使用流式细胞术检测各处理组细胞的凋亡率;④ 使用RT-PCR法检测各组K-RAS、ERK1、ERK2 mRNA的相对表达量.结果 ① 倒置显微镜下观察可见与对照组相比,除PD98059组外,其他各组细胞数量均减少,形态均发生凋亡变化,且随雷替曲塞浓度增高,变化程度增加;联合组细胞形态及数量变化较对应雷替曲塞组程度更深.② 细胞计数法显示各组细胞数均明显低于对照组的细胞数,且随着雷替曲塞浓度的增加,细胞数呈递减趋势;联合组与其他各组间均存在明显差异;③ 流式细胞仪检测显示和对照组相比,各组凋亡率均明显上升,在雷替曲塞组中存在浓度依赖性;联合组与其他各组的细胞凋亡率均存在差异;④ RT-PCR结果显示:与对照组相比,PD98059组、雷替曲塞组及联合组在K-RAS、ERK1、ERK2 mRNA的表达上均未表现出明显的差异.结论 ① 雷替曲塞对结肠癌细胞HT-29增殖及凋亡的影响呈浓度依赖性;② 雷替曲塞联合ERK抑制剂对HT-29细胞具有协同作用;③ ERK抑制剂不是在基因表达水平影响细胞的增殖及凋亡.

Effects of Raltitrexed and combination use of PD98059 on HT-29 cells

Objective To study the effects of Raltitrexed and combination use of PD98059 on HT-29 cell proliferation and apoptosis and explore their potential molecular mechanisms.Methods ① Inverted microscope was used to observe changes in cell morphology and counting of each group after treatment for 24 hours.② Cell counting method was used to detect cell quantity in each group.③ Flow cytometry instrument was used to test the apoptosis rate of every groups.④ RT-PCR method were used to detect each group′s K-RAS, ERK1, ERK2 mRNA expression.Results ① Each group had fewer cells and apoptotic changes except PD98059 group under inverted microscope,and the concentration of Raltitrexed higher,the change deeper.Changes in the joint group deeper than that of Raltitrexed group on cell quantity and form.② The result showed that the number of cells of each group were lower than control group,with higher concentration of raltitrexed, cell quantity decreased,and joint group had significant difference with other group exerted by cell counting method.③ Flow cytometry instrument testing showed that cell apoptosis of each group was increased compared with control group significantly, and Raltitrexed groups depended on concentra-tion, and joint group had significant difference with other group.④ There was no significant difference on K-RAS,ERK1,ERK2 mRNA expression in every groups by RT-PCR.Conclusion ① Raltitrexed exerts anti-proliferation and apoptosis-promoting effects on colon cancer cell HT-29 in a concentration dependent manner.② Raltitrexed combined with ERK inhibitor has synergistic effect on HT-29 cells.③ ERK inhibitor does not affect cell proliferation and apoptosis at gene expression levels.