miR-145对乳腺癌细胞及miR-100对卵巢癌细胞生物学特性的影响

发布时间：2018-02-04 06:26

本文关键词： 乳腺癌 miR-145 miR-100 迁移 TGF-β1 STAT3　出处：《江苏大学》2017年硕士论文　论文类型：学位论文

【摘要】：研究目的:微小RNA(mi RNA)是一种内源性的不能编码蛋白质的RNA。研究发现多种mi RNAs可以参与肿瘤的发生、发展进程并调节细胞生长、增殖、分化和凋亡。此外,报道发现miRNA的异常表达与肿瘤的发生、发展及转移有关。miR-145和mi R-100都是比较常见的mi RNAs,在多种肿瘤中异常表达,表现为促进或抑制肿瘤的进展。定量PCR检测发现miR-145在乳腺癌组织中的表达比在对应的癌旁组织中低,然后用miR-145 mimics和miR-145 inhibitor转染乳腺癌细胞,检测细胞在生长、迁移、增殖等方面的变化和对相关基因表达的影响,研究miR-145对乳腺癌细胞特性的影响,研究miR-145对乳腺癌的诊断或治疗的影响。此外,用miR-100 mimics和miR-100inhibitor转染卵巢癌细胞SKOV3,研究细胞生长、增殖以及迁移能力的变化,通过western blot实验检测miR-100处理后细胞的相关基因在蛋白水平表达的变化,研究miR-100对卵巢癌的诊断或治疗的影响。研究方法:(1)用定量PCR技术,检测在乳腺癌组织及对应的癌旁组织中mi R-145的表达水平,用软件分析miR-145在两种组织中的表达水平;(2)用顺转的方法将mir-145mimics和mir-145inhibitor转染到乳腺癌细胞中,观察细胞mcf-7和mda-mb-231在生长、迁移、增殖等方面的变化;并在卵巢癌细胞skov3中转染mir-100mimics和mir-100inhibitor,研究细胞生长、增殖及迁移能力的变化;(3)通过westernblot实验以及定量pcr实验检测mir-145处理后细胞的相关基因(转化生长因子,transforminggrowthfactor-β1,tgf-β1)在蛋白水平和mrna水平表达的变化;通过westernblot实验检测mir-100处理后细胞的相关基因(信号传导与转录激活因3,signaltransducerandactivatoroftranscription3,stat3)在蛋白水平表达的变化。研究结果:(1)通过定量pcr实验,发现mir-145在乳腺癌组织中的表达下降,经统计分析有统计学意义;(2)mir-145mimics抑制乳腺癌细胞的生长、迁移、增殖等能力,而mir-145inhibitor促进乳腺癌细胞的生长、迁移和增殖;(3)mir-100mimics抑制卵巢癌细胞skov3的生长、迁移、增殖等能力,而mir-100inhibitor促进卵巢癌细胞skov3的生长、迁移和增殖;(4)通过westernblot实验以及定量pcr实验检测mir-145处理后细胞的相关基因TGF-β1在蛋白水平和mRNA水平表达的变化,发现miR-145处理细胞后TGF-β1的表达在mRNA水平没有明显变化,在蛋白水平的表达降低;western blot实验检测miR-100处理后细胞SKOV3相关基因STAT3蛋白水平表达的变化,发现miR-100处理细胞后STAT3在蛋白水平的表达降低。结论:(1)miR-145在乳腺癌组织中低表达,miR-145表达降低的现象有希望成为乳腺癌诊断的新标准;(2)miR-145负向调控乳腺癌细胞的迁移和增殖,通过恢复病人mi R-145的表达,将有可能为乳腺癌的治疗提供帮助;(3)miR-100负向调控卵巢癌细胞的迁移和增殖,通过恢复病人mi R-100的表达,将有可能为卵巢癌的治疗提供帮助。
[Abstract]:Objective: small RNA(mi RNAs are endogenous RNAs that can not encode proteins. Many kinds of mi RNAs have been found to be involved in tumorigenesis. Development and regulation of cell growth, proliferation, differentiation and apoptosis. In addition, abnormal expression of miRNA and tumorigenesis have been reported. Development and metastasis related to. MiR-145 and miR-100 are more common in the expression of mi RNAs. they are abnormal in many kinds of tumors. Quantitative PCR analysis showed that the expression of miR-145 in breast cancer tissues was lower than that in corresponding paracancerous tissues. Then the breast cancer cells were transfected with miR-145 mimics and miR-145 inhibitor, and the growth and migration of the cells were detected. The effects of miR-145 on the characteristics of breast cancer cells and the effects of miR-145 on the diagnosis or treatment of breast cancer were studied. Ovarian cancer cell line SKOV3 was transfected with miR-100 mimics and miR-100inhibitor to study the changes of cell growth, proliferation and migration ability. Western blot assay was used to detect the expression of related genes in the cells treated with miR-100 at the protein level. To study the effect of miR-100 on the diagnosis or treatment of ovarian cancer. The expression of miR-145 was detected in breast cancer tissues and adjacent tissues, and the expression level of miR-145 was analyzed by software. (2) transfection of mir-145mimics and mir-145inhibitor into breast cancer cells by transposition. The changes of mcf-7 and mda-mb-231 in growth, migration and proliferation were observed. Mir-100mimics and mir-100 inhibitors were transfected into ovarian cancer skov3 to study the changes of cell growth, proliferation and migration ability. The related genes (transforming growth factor) of cells treated with mir-145 were detected by westernblot assay and quantitative pcr assay. The expression of transforming growth factor- 尾 1tgf- 尾 1 at protein level and mrna level; The related genes (signal transduction and transcriptional activation factor 3) of the cells treated with mir-100 were detected by westernblot assay. Signaltransducerandactivatoroftranscription3. The expression of stat3) at protein level. The results showed that the expression of mir-145 in breast cancer tissues was decreased by quantitative pcr assay, and there was statistical significance by statistical analysis. Mir-145inhibitor inhibited the growth, migration and proliferation of breast cancer cells, while mir-145inhibitor promoted the growth, migration and proliferation of breast cancer cells. The ability of growth, migration and proliferation of ovarian cancer cell line skov3 was inhibited by Mil 100 mimics. Mir-100inhibitor promoted the growth, migration and proliferation of ovarian cancer cell line skov3. (4) the expression of TGF- 尾 1 was detected by westernblot assay and quantitative pcr assay. The expression of TGF- 尾 1 was detected at the protein level and mRNA level after mir-145 treatment. It was found that the expression of TGF- 尾 1 did not change at the level of mRNA, but decreased at the level of protein after miR-145 treatment. Western blot assay was used to detect the expression of SKOV3 related gene STAT3 protein after miR-100 treatment. It was found that the expression of STAT3 in the cells treated with miR-100 was lower than that in the control group. Conclusion the expression of STAT3 is low in breast cancer. The low expression of miR-145 may become a new standard for the diagnosis of breast cancer. MiR-145 negatively regulates the migration and proliferation of breast cancer cells, which may be helpful for the treatment of breast cancer by restoring the expression of miR-145 in patients. MiR-100 negatively regulates the migration and proliferation of ovarian cancer cells, which may be helpful for the treatment of ovarian cancer by restoring the expression of miR-100 in patients.
【学位授予单位】：江苏大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R73-3

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