【摘要】：【目的】通過研究miR-506在宮頸癌組織和細胞中的表達水平及生物學功能，探討miR-506靶向調(diào)控Hedgehog信號通路中的Gli3基因的作用機制。 【方法（】1）用Real time PCR方法檢測50組配對的宮頸癌和癌旁組織中miR-506和Ki67的mRNA表達水平，比較miR-506在癌組織和癌旁組織中的表達差異并配對分析其表達趨勢；用Spearman相關(guān)性分析Ki67與miR-506表達水平之間的相關(guān)性；通過細胞增殖、細胞周期、細胞凋亡、caspase3/7活性檢測、紫杉醇和順鉑敏感性檢測及裸鼠皮下成瘤等實驗，過表達或干擾miR-506的表達后探討miR-506在宮頸癌中的生物學功能。（2）雙熒光素酶報告基因系統(tǒng)檢測miR-506對Gli3基因的調(diào)控作用；干擾或過表達Gli3基因，通過宮頸癌細胞增殖、細胞周期、細胞凋亡、caspase3/7活性、紫杉醇和順鉑藥物敏感性等實驗，明確能否部分再現(xiàn)或拮抗miR-506所誘導的生物學功能的改變。 【結(jié)果】（1）miR-506在宮頸癌組織中的表達水平明顯低于癌旁組織； Ki67與miR-506表達水平呈負相關(guān)；轉(zhuǎn)染miR-506的mimics后可抑制宮頸癌細胞的增殖、阻滯G1到S期的轉(zhuǎn)變、促進細胞凋亡、上調(diào)caspase3/7活性、增強紫杉醇和順鉑化療藥物敏感性（P＜0.05），干擾miR-506的表達能獲得與上述相反的結(jié)果；裸鼠皮下成瘤實驗亦證實miR-506能抑制宮頸癌細胞的生長。（2）雙熒光素酶報告基因系統(tǒng)結(jié)果顯示Gli3為miR-506的靶基因之一；沉默或過表達Gli3基因，，能部分再現(xiàn)或拮抗miR-506所誘導的上述生物學功能的改變。 【結(jié)論】miR-506在宮頸癌組織中表達水平低于癌旁組織，它通過靶向調(diào)控Gli3基因影響宮頸癌細胞的生物學功能，在宮頸癌的發(fā)病過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.
【學位授予單位】：上海交通大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R737.33

【參考文獻】

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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

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