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作 者:郭枫[1] 钟鸣[1] 杨乃林[1] 卞正乾[1] 赵刚[1]
机构地区:[1]上海交通大学医学院附属仁济医院普外科,上海200127
出 处:《现代生物医学进展》2014年第19期3684-3686,共3页Progress in Modern Biomedicine
摘 要:目的:检测CD133不同亚群大肠癌细胞HT-29的miR-429表达情况,探讨miR-429及CD133的表达与肿瘤的发生发展之间的关系。方法:采用荧光活化细胞分选法(FACS)分选出CD133不同亚群细胞,实时荧光定量PCR分别检测两组细胞miR-429的表达,合成miR-429寡核苷酸和阴性对照miRNA并分别转染CD133+和CD133-两个亚群细胞。再将细胞种植于非肥胖糖尿病/严重联合免疫缺陷(NOD/SCID)小鼠体内构建移植瘤模型,不同时间测量肿瘤体积和重量,RT-PCR及蛋白质印迹检测CD133+和CD133-两组肿瘤CD133mRNA和蛋白质表达。结果:血清检出CD133+细胞为67.9%,miR-429的表达量是CD133+细胞的(1.83±0.91)倍(P<0.05),CD133+比例与miR-429表达呈负相关(r=0.591,P<0.05);miR-429+/CD133+组的移植瘤体积及重量与对照组比较有统计学差异(P<0.05),且miR-429+/CD133+组成瘤时间较对照组晚约2周,但miR-429+/CD133+组的移植瘤CD133表达量低,与阴性对照组比较无明显差异(P>0.05)。结论:miR-429可能作为CD133的负性调控因子,具有抑制肿瘤生长的作用,但miR-429与CD133在肿瘤发生、发展过程中的作用机制有待进一步研究阐明。Objective: To detect the expressions of miR-429 in different CD133 subgroups on the HT-29 cells in the colorectal cancer tissues of which to explore the corelafion between the occurrence and progression of tumors. Methods: The different CD133 subgroup cells were sorted out by fluorescence activated cell sorting (FACS); the expression ofmiR-429 was detected by Real-time PCR; synthetic oligonucleotides of miR-429 and the negative control of microRNA were transfected in CD133+ and CD133+ subgroups; the non-obese diabetic/severe combined immunodeficiency mice (NOD/SCID) were planted the transfected cells to construct the transplanted tumor model; the tumor volume and weight were measured in different time; the expression of CD133 mRNA and protein in these two groups were detected by RT-PCR and western blotting. Results: The proportion of CD133+cells was 67.9%; the expression of miR-429 in CD133 cell was (1.83±0.91) times ofCD133+s(P〈0.05); the corelation offD133+ 's ratio and the expression ofmiR-429 was negative(r=0. 591, P〈0.05); there were statistically significant differences in the volume and weight of transplanted tumors in miR-429+/CD133+ between the two groups(P〈0.05); the tumorigenicity of miR-429+/CD133+ group was two weeks later than that of the control group, while the CD133 expression was lower with no significant difference than the negative control group(P〉0.05). Conclusion: It is suggested that the miR-429 might be a negative regulatory factor of CD133 which could inhibite the growth ofturnor. However, the mechanism of miR-429 and CD133 in process of tumor occurrence and development needs to be further studied and clarified.
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