本文關(guān)鍵詞：MiR-126調(diào)控cofilin-1與上皮性卵巢癌侵襲轉(zhuǎn)移關(guān)系的研究　出處：《浙江大學》2016年碩士論文　論文類型：學位論文

  更多相關(guān)文章： 卵巢癌 侵襲 miR-126 Cofilin-1

【摘要】：研究背景和目的：卵巢癌因其進展迅速,容易腹腔內(nèi)廣泛轉(zhuǎn)移,因此在確診時多數(shù)患者已處于晚期,目前已經(jīng)成為婦科致死率最高的腫瘤之一。卵巢癌細胞惡性生物學行為強,大部分患者接受標準化治療后2年左右出現(xiàn)腫瘤復發(fā)及耐藥,因此晚期卵巢癌患者5年生存率較低。近年來研究表明各種miRNA可通過作用于]mRNA轉(zhuǎn)錄后水平在腫瘤發(fā)生發(fā)展中起到抑癌或促癌作用。對于miR-126,有研究表明在乳腺癌、肺癌、前列腺癌等多種腫瘤發(fā)生發(fā)展中起著抑癌基因的作用,但其在卵巢癌中的作用尚不明確。絲切蛋白(Cofilin)是一種肌動蛋白結(jié)合蛋白,在細胞骨架重塑和移動調(diào)節(jié)上起著重要作用,而卵巢癌等惡性腫瘤細胞的分裂、遷移及特異性結(jié)構(gòu)依賴于細胞骨架的動力學調(diào)節(jié)。為此本研究擬通過體外培養(yǎng)SKOV3卵巢癌細胞株并轉(zhuǎn)染不同miR-126基因片段,比較miR-126與Cofilia-1表達之間的關(guān)系,以此研究miR-126是否可通過調(diào)控conflin-1而參與卵巢癌細胞的侵襲轉(zhuǎn)移的調(diào)節(jié)。研究材料和方法：本研究將人卵巢漿液性囊腺癌SKOV3細胞株進行體外培養(yǎng),miR-126通過慢病毒的包裝后轉(zhuǎn)染人卵巢漿液性囊腺癌SKOV3細胞系,上述卵巢癌細胞分成四組,分別為：miR-126過表達組(SKOV3+LV3-has-miR-126組)、miR-126抑制表達組(SKOV3+LV3-has-miR-126 inhibitor組)、轉(zhuǎn)染陰性對照組(SKOV3+LV3NC組)和空白對照組(SKOV3組),實驗采用Transwell侵襲實驗觀察各組細胞的侵襲能力,通過免疫熒光法觀察cofilin-1表達水平的差異。結(jié)果：1.Transwell侵襲實驗的結(jié)論表明miR-126過表達組(SKOV3+LV3-has-miR-126組SKOV3細胞侵犯基質(zhì)膠的穿膜細胞數(shù)明顯少于miR-126抑制表達組(SKOV3+LV3-has-miR-126 inhibitor組)(P0.05),同時也明顯少于轉(zhuǎn)染陰性對照組(SKOV3+LV3NC組)和空白對照組(SKOV3組)的穿膜細胞數(shù)(P均0.05)。2.免疫熒光檢測結(jié)果顯示,LV3-has-miR-126組細胞Cofilin-1表達明顯低于miR-126抑制表達組(SKOV3+LV3-has-miR-126 inhibitor組)、轉(zhuǎn)染陰性對照組(SKOV3+LV3NC組)和空白對照組(SKOV3組),而LV3-has-miR-126 inhibitor組Cofilin-1表達明顯高于轉(zhuǎn)染陰性對照組(SKOV3+LV3NC組)和空白對照組(SKOV3組)。結(jié)論：1. miR-126過表達可以減弱SKOV3細胞侵襲能力,提示miR-126過表達可能抑制卵巢癌細胞的侵襲轉(zhuǎn)移。2.轉(zhuǎn)染miR-126的SKOV3細胞cofilin-1表達明顯減少,提示miR-126可能通過抑制cofil in-1表達,進而影響細胞骨架而抑制卵巢癌細胞的侵襲轉(zhuǎn)移。
[Abstract]:Background and objective: ovarian cancer is in advanced stage because of its rapid progression and easy metastasis in abdominal cavity. Ovarian cancer cells have strong malignant biological behavior, most of the patients received standardized treatment about 2 years after the tumor recurrence and drug resistance. Therefore, the 5-year survival rate of patients with advanced ovarian cancer is low. In recent years, studies have shown that various kinds of miRNA can inhibit or promote cancer in tumorigenesis and development by acting on] mRNA posttranscriptional level. -126. Studies have shown that breast cancer, lung cancer, prostate cancer and other tumors play a role in the development of tumor suppressor genes. However, its role in ovarian cancer is unclear. Cofilin is an actin binding protein, which plays an important role in cytoskeletal remodeling and movement regulation. And the division of malignant tumor cells such as ovarian cancer. Migration and specific structure depend on the dynamics of cytoskeleton. This study aims to culture SKOV3 ovarian cancer cell lines in vitro and transfect different miR-126 gene fragments. To compare the relationship between miR-126 and Cofilia-1 expression. To study whether miR-126 is involved in the regulation of invasion and metastasis of ovarian cancer cells by regulating conflin-1. Materials and methods:. In this study, human ovarian serous cystadenocarcinoma cell line SKOV3 was cultured in vitro. MiR-126 was transfected into human ovarian serous cystadenocarcinoma SKOV3 cell line after packaging of lentivirus. The ovarian cancer cells were divided into four groups. SKOV3 LV3-has-miR-126 group was the overexpressed group of 10% miR-126 (P < 0.05). MiR-126 inhibited expression of SKOV3 LV3-has-miR-126 inhibitor. SKOV3 LV3NC (negative control group) and SKOV3 (blank control group) were transfected. Transwell invasion assay was used to observe the invasiveness of the cells in each group. The expression of cofilin-1 was observed by immunofluorescence method. Results 1. The results of Transwell invasion experiment showed that the miR-126 overexpression group (. The number of SKOV3 cells invading matrix glue in SKOV3 LV3-has-miR-126 group was significantly less than that in miR-126 inhibited expression group (2). SKOV3 LV3-has-miR-126 inhibitor group (P0.05). At the same time, the number of perforating cells in SKOV3 LV3NC group (negative transfection group) and SKOV3 group (blank control group) were significantly lower than that in SKOV3 group. The expression of Cofilin-1 in LV3-has-miR-126 group was significantly lower than that in miR-126 group (. SKOV3 LV3-has-miR-126 inhibitor group. Transfection negative control group (SKOV3 LV3NC group) and blank control group (SKOV3 group). The expression of Cofilin-1 in LV3-has-miR-126 inhibitor group was significantly higher than that in negative control group (SKOV3 LV3NC group) and blank control group (P < 0.05). Conclusion: 1. Overexpression of miR-126 can attenuate the invasiveness of SKOV3 cells. These results suggest that the overexpression of miR-126 may inhibit the invasion and metastasis of ovarian cancer cells. 2. The expression of cofilin-1 in SKOV3 cells transfected with miR-126 was significantly decreased. The results suggest that miR-126 inhibits the invasion and metastasis of ovarian cancer cells by inhibiting the expression of cofil in-1 and thus affecting the cytoskeleton.
【學位授予單位】：浙江大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R737.31

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本文編號：1357441