miR-506靶向调控Gli3基因在宫颈癌中的作用及机制研究
发布时间：2018-10-21 13:21
【摘要】：【目的】通过研究miR-506在宫颈癌组织和细胞中的表达水平及生物学功能，探讨miR-506靶向调控Hedgehog信号通路中的Gli3基因的作用机制。 【方法（】1）用Real time PCR方法检测50组配对的宫颈癌和癌旁组织中miR-506和Ki67的mRNA表达水平，比较miR-506在癌组织和癌旁组织中的表达差异并配对分析其表达趋势；用Spearman相关性分析Ki67与miR-506表达水平之间的相关性；通过细胞增殖、细胞周期、细胞凋亡、caspase3/7活性检测、紫杉醇和顺铂敏感性检测及裸鼠皮下成瘤等实验，过表达或干扰miR-506的表达后探讨miR-506在宫颈癌中的生物学功能。（2）双荧光素酶报告基因系统检测miR-506对Gli3基因的调控作用；干扰或过表达Gli3基因，通过宫颈癌细胞增殖、细胞周期、细胞凋亡、caspase3/7活性、紫杉醇和顺铂药物敏感性等实验，明确能否部分再现或拮抗miR-506所诱导的生物学功能的改变。 【结果】（1）miR-506在宫颈癌组织中的表达水平明显低于癌旁组织； Ki67与miR-506表达水平呈负相关；转染miR-506的mimics后可抑制宫颈癌细胞的增殖、阻滞G1到S期的转变、促进细胞凋亡、上调caspase3/7活性、增强紫杉醇和顺铂化疗药物敏感性（P＜0.05），干扰miR-506的表达能获得与上述相反的结果；裸鼠皮下成瘤实验亦证实miR-506能抑制宫颈癌细胞的生长。（2）双荧光素酶报告基因系统结果显示Gli3为miR-506的靶基因之一；沉默或过表达Gli3基因，，能部分再现或拮抗miR-506所诱导的上述生物学功能的改变。 【结论】miR-506在宫颈癌组织中表达水平低于癌旁组织，它通过靶向调控Gli3基因影响宫颈癌细胞的生物学功能，在宫颈癌的发病过程中可能起重要作用。
[Abstract]:[objective] to study the expression level and biological function of miR-506 in cervical cancer tissues and cells. To investigate the mechanism of miR-506 targeting the regulation of Gli3 gene in Hedgehog signaling pathway. [methods] 1) the mRNA expression of miR-506 and Ki67 in 50 pairs of cervical cancer and adjacent tissues were detected by Real time PCR method. To compare the difference of miR-506 expression in cancer tissues and adjacent tissues and to analyze the expression trend of Ki67 by paired analysis; to analyze the correlation between Ki67 and miR-506 expression level by Spearman correlation; to detect the activity of caspase3/7 by cell proliferation, cell cycle, apoptosis and caspase3/7 activity. The biological function of miR-506 in cervical cancer was investigated after overexpression or interference with miR-506 expression. (2) double luciferase reporter gene system was used to detect the regulatory effect of miR-506 on Gli3 gene. Interference or overexpression of Gli3 gene was detected by cell proliferation, cell cycle, apoptosis, caspase3/7 activity, paclitaxel and cisplatin susceptibility. [results] [results] (1) the expression of miR-506 in cervical carcinoma was significantly lower than that in paracancerous tissues, and the expression of Ki67 was negatively correlated with that of miR-506. Mimics transfected with miR-506 inhibited the proliferation of cervical cancer cells, blocked the transition from G1 to S, promoted apoptosis, upregulated the activity of caspase3/7, enhanced the chemosensitivity of paclitaxel and cisplatin (P < 0. 05), and interfered with the expression of miR-506. Subcutaneous tumorigenesis in nude mice also confirmed that miR-506 could inhibit the growth of cervical cancer cells. (2) double luciferase reporter gene system showed that Gli3 was one of the target genes of miR-506, and the Gli3 gene was silenced or overexpressed. [conclusion] the expression level of miR-506 in cervical cancer tissues is lower than that in adjacent tissues, and it can affect the biological function of cervical cancer cells by targeting Gli3 gene, which can partially reproduce or antagonize the above changes of biological function induced by miR-506. [conclusion] the expression level of miR-506 in cervical cancer tissues is lower than that in adjacent tissues. It may play an important role in the pathogenesis of cervical cancer.
【学位授予单位】：上海交通大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R737.33
本文编号：2285203
[Abstract]:[objective] to study the expression level and biological function of miR-506 in cervical cancer tissues and cells. To investigate the mechanism of miR-506 targeting the regulation of Gli3 gene in Hedgehog signaling pathway. [methods] 1) the mRNA expression of miR-506 and Ki67 in 50 pairs of cervical cancer and adjacent tissues were detected by Real time PCR method. To compare the difference of miR-506 expression in cancer tissues and adjacent tissues and to analyze the expression trend of Ki67 by paired analysis; to analyze the correlation between Ki67 and miR-506 expression level by Spearman correlation; to detect the activity of caspase3/7 by cell proliferation, cell cycle, apoptosis and caspase3/7 activity. The biological function of miR-506 in cervical cancer was investigated after overexpression or interference with miR-506 expression. (2) double luciferase reporter gene system was used to detect the regulatory effect of miR-506 on Gli3 gene. Interference or overexpression of Gli3 gene was detected by cell proliferation, cell cycle, apoptosis, caspase3/7 activity, paclitaxel and cisplatin susceptibility. [results] [results] (1) the expression of miR-506 in cervical carcinoma was significantly lower than that in paracancerous tissues, and the expression of Ki67 was negatively correlated with that of miR-506. Mimics transfected with miR-506 inhibited the proliferation of cervical cancer cells, blocked the transition from G1 to S, promoted apoptosis, upregulated the activity of caspase3/7, enhanced the chemosensitivity of paclitaxel and cisplatin (P < 0. 05), and interfered with the expression of miR-506. Subcutaneous tumorigenesis in nude mice also confirmed that miR-506 could inhibit the growth of cervical cancer cells. (2) double luciferase reporter gene system showed that Gli3 was one of the target genes of miR-506, and the Gli3 gene was silenced or overexpressed. [conclusion] the expression level of miR-506 in cervical cancer tissues is lower than that in adjacent tissues, and it can affect the biological function of cervical cancer cells by targeting Gli3 gene, which can partially reproduce or antagonize the above changes of biological function induced by miR-506. [conclusion] the expression level of miR-506 in cervical cancer tissues is lower than that in adjacent tissues. It may play an important role in the pathogenesis of cervical cancer.
【学位授予单位】：上海交通大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R737.33
【参考文献】
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1 Seow Chong Lee;Hwee Tong Tan;Maxey Ching Ming Chung;;Prognostic biomarkers for prediction of recurrence of hepatocellular carcinoma:Current status and future prospects[J];World Journal of Gastroenterology;2014年12期
本文编号：2285203
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