微RNA-21表达下调可抑制人结肠癌SW620细胞在裸鼠体内的生长

发布时间：2018-02-23 18:39

本文关键词： 结肠肿瘤微RNAs 细胞凋亡异种移植模型抗肿瘤试验小鼠近交BALB/c　出处：《肿瘤》2017年02期 　论文类型：期刊论文

【摘要】：目的 :探讨反义寡核苷酸(antisense oligonucleotides,ASOs)下调微RNA-21(micro RNA-21,mi R-21)表达对高侵袭性人结肠癌SW620细胞在裸鼠体内生长的影响。方法 :首先建立人结肠癌SW620细胞的裸鼠荷瘤模型,并于肿瘤局部分别注射p-mi R-21-ASOs重组质粒或p-Cont对照质粒,然后观察裸鼠体内肿瘤的生长变化。末次注射5 d后,处死裸鼠,剥取肿瘤组织,测量各组肿瘤组织的大小和质量。采用HE染色法观察肿瘤组织形态学变化;TUNEL法检测肿瘤局部细胞凋亡情况;免疫组织荧光法检测肿瘤组织中增殖核抗原Ki-67蛋白的表达变化;实时荧光定量PCR法检测肿瘤组织中mi R-21成熟体的表达,以及细胞周期蛋白依赖性激酶(cyclin-dependent kinase,CDK)2、CDK3、CDK4和CDK6的表达变化;蛋白质印迹法检测磷酸化蛋白激酶B(phosphorylated protein kinase B,p-PKB,又称为p-Akt)和磷酸化细胞外信号调节激酶(phosphorylated extracellular regulated protein kinase,p-ERK)的表达水平。结果 :与p-Cont对照组相比,p-mi R-21-ASOs质粒转染组的肿瘤生长缓慢(P0.05),剥离肿瘤的质量明显减少(P0.01),肿瘤组织中可见大片坏死区域,而且肿瘤细胞中mi R-21成熟体的表达水平明显下调(P0.01)。mi R-21表达下调后,肿瘤组织中Ki-67的表达量明显减少(P0.01),而相对凋亡细胞数明显增多(P0.05);并且,细胞周期相关分子CDK2、CDK3、CDK4和CDK6的表达均明显下调(P值均0.001),Akt和ERK蛋白的磷酸化水平也均明显下调(P值均0.05)。结论 :ASOs下调mi R-21表达可显著抑制人高侵袭性结肠癌细胞在裸鼠体内的生长,提示mi R-21可作为结肠癌基因治疗的一个潜在新靶标。
[Abstract]:Objective: To investigate the effect of antisense oligonucleotides (antisense oligonucleotides, ASOs RNA-21 (micro) by micro RNA-21, MI R-21) on the expression of highly invasive human colon cancer SW620 cells in nude mice. Methods: the growth of nude mice model established human colon cancer cell line SW620, and to the local tumor were injected p-mi R-21-ASOs recombinant plasmid p-Cont or control plasmid, then we observed the growth of nude mice tumor. 5 d after the last injection, mice were sacrificed, stripping of tumor tissue, size and quality were measured in tumor tissue. HE was used to observe the morphological changes of tumor tissue staining; apoptosis detected by TUNEL tumor cells; expression of proliferating cell nuclear antigen Ki-67 immunofluorescence detection of protein expression in tumor tissue; tumor tissues were detected by real time fluorescent quantitative PCR MI in mature R-21, and cyclin dependent kinase Enzyme (cyclin-dependent kinase, CDK 2, CDK3), the expression of CDK4 and CDK6; detection of phosphorylated protein kinase B protein blotting (phosphorylated protein kinase B p-PKB, also known as p-Akt) and phosphorylated extracellular signal regulated kinase (phosphorylated extracellular regulated protein kinase, p-ERK) expression level. Results: compared with the p-Cont control group, p-mi transfection of R-21-ASOs plasmid group (P0.05), slow tumor growth of tumor quality significantly decreased (P0.01), observed in tumor tissue necrosis region, and the expression level of MI in tumor cells of mature R-21 (P0.01).Mi was downregulated after downregulation of R-21 expression of Ki-67 in tumor tissues decreased (P0.01), and the relative number of apoptotic cells increased significantly (P0.05); and the cell cycle related molecules CDK2, CDK3, CDK4 and CDK6 expression were significantly reduced (P 0.001), Akt and ERK. The level of phosphorylation of white was also significantly down regulated (P = 0.05). Conclusion: ASOs downregulation of MI R-21 expression can significantly inhibit the growth of highly invasive colon cancer cells in nude mice, suggesting that MI R-21 can be used as a potential new target for gene therapy of colon cancer.

【作者单位】：遵义医学院免疫学教研室&贵州省生物治疗人才基地;遵义医学院贵州省普通高等学校特色药物肿瘤防治重点实验室;遵义医学院医学物理学教研室;贵州医科大学组织工程与干细胞实验中心;
【基金】：贵州省应用基础研究计划重大专项(编号:黔科合J重大字[2015]2003) 贵州省高层次创新型人才培养项目(编号:黔科合人才[2016]4031号) 遵义医学院优秀青年人才计划(编号:15ZY-001)~~
【分类号】：R735.35

【相似文献】