本文選題：長鏈非編碼RNA + linc-UBC1�。� 參考：《南方醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的現(xiàn)已有大量的研究證明了長鏈非編碼RNA(lncRNA)對惡性腫瘤發(fā)生發(fā)展的調(diào)控作用。據(jù)研究報道長鏈非編碼RNA linc-UBC1可通過結(jié)合PRC2而影響膀胱癌預(yù)后。本研究擬通過檢測linc-UBC1在結(jié)直腸癌(CRC)中的表達(dá),探索linc-UBC1對CRC細(xì)胞生物學(xué)功能的影響,初步揭示linc-UBC1對CRC的作用,為今后CRC的診斷和治療提供新的思路。方法1.收集96例結(jié)直腸癌患者的手術(shù)標(biāo)本(包含癌組織和對應(yīng)癌旁組織)和患者的臨床病理資料,結(jié)直腸癌細(xì)胞系(SW480、SW620、HCT116和HT29)以及正常結(jié)腸上皮細(xì)胞NCM460由南方醫(yī)科大學(xué)捐贈。2.通過實時熒光定量聚合酶鏈反應(yīng)技術(shù)(RT-qPCR)檢測結(jié)直腸癌組織和對應(yīng)癌旁組織linc-UBC1的表達(dá)量,并分析其與臨床病理資料的關(guān)系,采用Kaplan—Meier法分析linc-UBC1與生存資料的關(guān)系。3.通過RT-qPCR檢測4種結(jié)直腸癌細(xì)胞系及正常結(jié)腸上皮細(xì)胞NCM460中l(wèi)inc-UBC1的表達(dá)量,選取linc-UBC1表達(dá)量最高的結(jié)直腸癌細(xì)胞系SW620,通過脂質(zhì)體轉(zhuǎn)染技術(shù)干擾SW620細(xì)胞系中的linc-UBC1基因,并使用RT-qPCR檢測其干擾效率。4.運用CCK8、流式細(xì)胞術(shù)及Transwell實驗檢測linc-UBC1基因干擾后的SW620細(xì)胞系的增殖能力、凋亡率及周期、遷移及侵襲能力的變化。5.運用western blot實驗檢測linc-UBC1基因干擾后的SW620細(xì)胞系的凋亡相關(guān)蛋白(cleaved caspase-3、cleaved caspase-9 和 Bcl-2)表達(dá)量的變化。結(jié)果1.linc-UBC1在結(jié)直腸癌組織中的相對表達(dá)量為6.55±6.60,遠(yuǎn)高于癌旁組織的相對表達(dá)量1.67±1.54,兩者比較差異有統(tǒng)計學(xué)意義(t=8.32,P0.001)。2.通過分析linc-UBC1的表達(dá)量與臨床病理的關(guān)系得出:linc-UBC1的表達(dá)量與結(jié)直腸癌的TNM分期、腫瘤的大小、浸潤的深度及淋巴結(jié)轉(zhuǎn)移相關(guān);差異有統(tǒng)計學(xué)意義(P0.05)。而與患者的性別、年齡、腫瘤的分化程度、腫瘤位置和遠(yuǎn)處轉(zhuǎn)移無關(guān)(P0.05)。生存分析結(jié)果顯示:linc-UBC1高表達(dá)組的生存時間明顯小于linc-UBC1低表達(dá)組,差異有統(tǒng)計學(xué)意義(P0.01)。多因素COX回歸分析表明:linc-UBC1的表達(dá)水平可作為CRC患者的獨立預(yù)后因子(Hazard ratio:2.434,95%CI:1.093-5.420,P=0.029)。3.與 NCM460 相比,SW620、HCT116 和 HT29 中 linc-UBC1 表達(dá)水平均明顯升高(P0.01),而在SW480細(xì)胞中無顯著差異(P0.05),其中SW620的linc-UBC1表達(dá)量最高。4.與陰性對照組相比,linc-UBC1干擾組的SW620的增殖、遷移及侵襲能力降低,凋亡率升高及周期受到抑制(P0.05)。5.與陰性對照組相比,linc-UBC1干擾組的SW620的凋亡相關(guān)蛋白cleaved caspase-3和cleavedcaspase-9的表達(dá)量顯著上升,Bcl-2的表達(dá)量顯著下降,差異有統(tǒng)計學(xué)意義(P0.05)。結(jié)論linc-UBC1在CRC組織中的表達(dá)量顯著升高,且linc-UBC1的表達(dá)量與CRC的TNM分期、腫瘤的大小、浸潤的深度、淋巴結(jié)轉(zhuǎn)移及不良預(yù)后正相關(guān),并且linc-UBC1可以促進CRC細(xì)胞的增殖、遷移及侵襲。因此,linc-UBC1有可能成為判斷CRC患者預(yù)后和發(fā)展的新的腫瘤標(biāo)記物。
[Abstract]:Objective A large number of studies have demonstrated the regulatory effect of long chain noncoding RNAs (LNRNAs) on the carcinogenesis and development of malignant tumors.It has been reported that long chain non-coding RNA linc-UBC1 can affect the prognosis of bladder cancer by binding PRC2.This study was designed to explore the effect of linc-UBC1 on the biological function of CRC cells by detecting the expression of linc-UBC1 in colorectal carcinoma, and to reveal the effect of linc-UBC1 on CRC, and to provide a new idea for the diagnosis and treatment of CRC in the future.Method 1.The clinical and pathological data of 96 patients with colorectal cancer (including cancer tissues and adjacent tissues) and the clinicopathological data were collected. The colorectal cancer cell lines SW480, SW620, HCT116 and HT29, as well as normal colonic epithelial cells NCM460, were donated by Southern Medical University.Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of linc-UBC1 in colorectal cancer tissues and adjacent tissues, and the relationship between linc-UBC1 and survival data was analyzed by Kaplan-Meier method.The expression of linc-UBC1 in NCM460 of four colorectal cancer cell lines and normal colonic epithelial cells was detected by RT-qPCR. The cell line SW620 with the highest linc-UBC1 expression was selected, and the linc-UBC1 gene in SW620 cell line was interfered with by liposome transfection technique.And use RT-qPCR to detect its interference efficiency. 4. 4.CCK8, flow cytometry and Transwell assay were used to detect the changes of proliferation, apoptosis rate and cycle, migration and invasion ability of SW620 cell lines after linc-UBC1 gene interference.Western blot assay was used to detect the expression of apoptosis-related proteins (caspase-9 and Bcl-2) in SW620 cell lines after linc-UBC1 gene interference.Results the relative expression of 1.linc-UBC1 in colorectal cancer was 6.55 鹵6.60, which was significantly higher than that in paracancerous tissues (1.67 鹵1.54).By analyzing the relationship between the expression of linc-UBC1 and clinicopathology, it was concluded that the expression of linc-UBC1 was correlated with the TNM stage, tumor size, depth of invasion and lymph node metastasis of colorectal cancer, and the difference was statistically significant (P 0.05).There was no correlation between sex, age, tumor differentiation, tumor location and distant metastasis (P 0.05).Survival analysis showed that the survival time of the high expression group of linc-UBC1 was significantly shorter than that of the low expression group of linc-UBC1, and the difference was statistically significant (P 0.01).Multivariate COX regression analysis showed that the expression level of BC1 in CRC patients could be regarded as the independent prognostic factor of CRC patients.Compared with NCM460, the expression of linc-UBC1 in HCT116 and HT29 was significantly higher than that in NCM460, but there was no significant difference in SW480 cells. The linc-UBC1 expression of SW620 was the highest. 4.Compared with the negative control group, the SW620 proliferation, migration and invasion ability of the control group were decreased, the apoptosis rate was increased and the cycle was inhibited by P0.05. 5.Compared with the negative control group, the expression of apoptosis-related protein cleaved caspase-3 and cleavedcaspase-9 in the SW620 interference group was significantly increased, and the expression of Bcl-2 was significantly decreased in the control group, and the difference was statistically significant (P 0.05).Conclusion the expression of linc-UBC1 in CRC tissues was significantly increased, and the expression of linc-UBC1 was positively correlated with the TNM stage of CRC, tumor size, depth of invasion, lymph node metastasis and poor prognosis. Linc-UBC1 could promote the proliferation, migration and invasion of CRC cells.Therefore, linc-UBC1 may be a new tumor marker for the prognosis and development of CRC patients.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R735.3
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本文編號：1769346