摘要
目的研究PRR13基因表达量与结直肠癌细胞对于奥沙利铂药物敏感性的影响以及在奥沙利铂药物作用下PRR13基因表达高低对结直肠癌细胞凋亡率的影响。方法在美国模式菌种收集中心(ATCC)常用结直肠癌细胞系中筛选出2株结直肠癌细胞系(SW620,SW480),通过慢病毒载体进行细胞转染构建PRR13沉默结直肠癌细胞模型,采用MTT法进行奥沙利铂药物敏感性实验,本研究将稳定转染包含沉默PRR13基因序列shRNA慢病毒的结直肠细胞模型定义为沉默组,将稳定转染慢病毒空载体的结直肠细胞模型定义为对照组。PRR13基因沉默后,奥沙利铂的半抑制浓度(IC50)变化,然后使用双染法流式细胞仪凋亡实验,比较组间PRR13表达量及与结直肠癌细胞凋亡率的关系。计量资料以均数±标准差(Mean±SD)表示,组间比较采用独立样本t检验;计数资料以百分比(%)表示,组间比较采用χ^2检验。结果相同细胞模型IC50随时间的增加而显著降低,相同时间点,沉默组IC50明显低于对照组;在相同浓度作用的同类细胞模型中,沉默组细胞凋亡率明显高于对照组[SW620沉默组(22.50%)比SW620对照组(11.35%),SW480沉默组(13.63%)比SW480对照组(4.59%)],沉默组活细胞比率明显低于对照组细胞模型[SW620沉默组(76.0%)比SW620对照组(87.2%),SW480沉默组(74.5%)比SW480对照组(89.3%)]。结论沉默PRR13基因可以降低细胞对于奥沙利铂药物的敏感性,降低奥沙利铂药物耐药性,化学治疗效果更显著。
Objective To study the effect of PRRI3 expression and the sensitivity of colorectal cancer cells to oxaliplatin,as well as the efect of PRRI3 expression on the apoptosis rate of colorectal cancer cells under the action of oxaliplatin.Methods Two colorectal cancer cell lines(SW620,SW480)were screened from the colorectal cancer cell lines commonly used by ATCC.Lentiviral vectors were used for cell transfection to construct a PRRI3-silent colorectal cancer cell model.MTT oxaliplatin drug sensitivity experiments were conducted and compared.In this study,a colorectal cell model stably transected with shRNA lentivirus pontaining the silenced PRRI3 sequence was defined as the silent group,and a colorectal cell model stably transfected with empty vector of lentivirus was defined as the control group.After silencing the PRRI3,the colorectal cancer cells changed the ICs0 of oxaliplatin,and then used a double staining flow cytomnetry apoptosis experiment to compare the expression of PRRI3 and the rate of apoptosis of colorectal cancer cells under the action of oxaliplatin relationship.The measurement data were expressed as mean±standard deviation(Mean±SD),and the comparison between groups used independent sample t test.The count data were expressed as percentage(%),and the comparison between groups used chi-square test.Results The IC3 of the same cell model decreased significantly with time.At the same time point,the IC of the silent group was significantly lower than that of the control group.In the same cell model with the same concentration,the apoptosis rate of the silent group was significantly higher than that of the control group(SW620 silent group 22.5%us SW620 control group 11.35%,SW480 silent group 13.63%a1s SW480 control group 4.59%),the ratio of viable cells was significantly lower than that of the control cell model(SW620 silent group 76.0%us SW620 control group 87.2%,SW480 silent group 74.5%us SW480 control group 89.3%).Conclusion Sileneing the PRRI3 can reduce the sensitivity of cells to oxaliplatin and red
作者
周晓娜
王今
金岚
张忠涛
Zhou Xiaona;Wang Jin;Jin Lan;Zhang Zhongtao(Department of General Surgery,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China)
出处
《国际外科学杂志》
2020年第7期464-470,F0004,共8页
International Journal of Surgery
基金
国家自然科学基金(81172318)。
