摘要
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。
Objective To study the effects of Raltitrexed and combination use of PD98059 on HT-29 cell prolifera- tion 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. Re- sults （1） Each group had fewer cells and apoptotie 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 Rahitrexed 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 Rahitrexed 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） Rahitrexed exerts anti-proliferation and apoptosis-promoting effects on colon cancer cell HT-29 in a concentration dependent manner. （2） Rahitrexed combined with ERK inhibitor has synergistic effect on HT-29 cells. （3） ERK inhibitor does not affect cell prolifera- tion and apoptosis at gene expression levels.
出处
《安徽医科大学学报》
CAS
北大核心
2017年第5期692-697,共6页
Acta Universitatis Medicinalis Anhui
基金
安徽省高等学校自然科学研究项目(编号:KJ2015A334)
