發(fā)布時間：2019-04-15 22:34

【摘要】：大腸癌(colorectal cancer,CRC)是最常見的消化系統(tǒng)惡性腫瘤之一,在全球范圍內(nèi)位居女性惡性腫瘤發(fā)病率的第2位,男性惡性腫瘤發(fā)病率的第3位。大腸癌為第5位惡性腫瘤致死病種,患者的生存和預(yù)后與大腸癌的首次診斷時間,治療方法以及對化療的敏感性等密切相關(guān)。因此,及時有效地診斷和治療大腸癌患者具有重要意義。微小RNA(micro RNA,mi RNA)是一種單鏈非編碼小分子RNA,長度為18~25個核苷酸,它可通過與靶m RNA 3’末端非翻譯區(qū)完全或不完全匹配結(jié)而誘導(dǎo)靶m RNA降解或阻遏其翻譯,從而在轉(zhuǎn)錄后水平沉默基因的表達(dá),進(jìn)而發(fā)揮其功能。目前,已在哺乳動物中發(fā)現(xiàn)超過1000種,經(jīng)研究確認(rèn)的mi RNA有533種。進(jìn)而使人們對基因表達(dá)的調(diào)控有了新的認(rèn)識。mi RNA在多種生物學(xué)過程,如細(xì)胞生長、增殖、分化和凋亡等發(fā)揮著重要的作用。mi RNA涉及多種疾病,包括心血管系統(tǒng)、神經(jīng)系統(tǒng)、免疫系統(tǒng)疾病和癌癥。mi RNA在多種惡性腫瘤中異常表達(dá),被證明參與腫瘤的發(fā)生、進(jìn)展、侵襲、轉(zhuǎn)移及血管生成等。在大腸癌中,mi RNA的異常表達(dá)與腫瘤的大小、分期和預(yù)后等均明顯相關(guān),并可能為大腸癌的早期診斷提供新的分子標(biāo)志物。微小RNA(mi RNA)可以作為腫瘤抑制或致癌基因。研究表明mi RNA-141水平在大腸癌組織樣本中上調(diào)。實驗觀察到很多mi R-14的可能靶目標(biāo),包括腫瘤抑制基因MAP2K4。本研究的目的是探討mi R-141促進(jìn)大腸癌增殖的作用。方法采用RT-PCR檢測20例確診結(jié)腸腺癌及其配對癌周正常組織中mi R-141以及MAP2K4的m RNA表達(dá)水平;分析mi R-141與TNM分期的相關(guān)性,利用mi R-141 inhibitor或mi R-141 mimics分析抑制或者增加細(xì)胞中mi R-141表達(dá)對結(jié)腸腺癌細(xì)胞系增殖的影響,利用si RNA抑制MAP2K4表達(dá),明確MAP2K4蛋白在mi R-141引起的大腸癌細(xì)胞增殖中的作用。免疫組化染色檢測大腸癌組織及癌周正常組織MAP2K4蛋白表達(dá)。Western blot檢測大腸癌細(xì)胞中MAP2K4蛋白水平。利用MTT法檢測大腸癌細(xì)胞增殖情況。結(jié)果miR-141在TNMⅢ+Ⅳ期的mi R-141 m RNA表達(dá)水平顯著高于Ⅰ+Ⅱ期。與癌旁正常組織相比,20例大腸癌臨床樣品中的mi R-141水平均顯著上調(diào)。與此相反,MAP2K4的表達(dá)水平均顯著下降。通過MTT細(xì)胞增殖測定實驗,過度表達(dá)mi R-141通過抑制MAP2K4活性導(dǎo)致大腸癌細(xì)胞增殖。結(jié)論miR-141與TNM分期相關(guān),mi R-141上調(diào)與大腸癌細(xì)胞增殖有關(guān),mi R-141是通過抑制腫瘤抑制基因MAP2K4而促進(jìn)癌細(xì)胞的生長。針對mi R-141的策略可以提供一個有效治療大腸癌的方法。
[Abstract]:Colorectal cancer (colorectal cancer,CRC) is one of the most common malignant tumors of digestive system. It is the second most common malignant tumor in female and the third in male. Colorectal cancer is the fifth leading cause of death from malignant tumors. The survival and prognosis of the patients are closely related to the first diagnosis time, treatment methods and sensitivity to chemotherapy of colorectal cancer. Therefore, timely and effective diagnosis and treatment of colorectal cancer patients have important significance. Small RNA (micro RNA,mi RNA) is a single-stranded non-coding small molecule with a length of 18 渭 25 nucleotides. It can induce the degradation of target m-RNA or inhibit its translation by completely or incompletely matching the untranslated region at the 3 'end of the target m RNA. Thus silencing the expression of the gene at post-transcriptional level, and then play its role. At present, more than 1000 species of mi RNA have been found in mammals and 533 species have been identified. Mi RNA plays an important role in many biological processes, such as cell growth, proliferation, differentiation and apoptosis. Mi RNA involves many diseases, including cardiovascular system, nervous system, and so on. The abnormal expression of. Mi RNA in many kinds of malignant tumors has been proved to be involved in tumorigenesis, progression, invasion, metastasis, angiogenesis and so on. The abnormal expression of, mi RNA is significantly correlated with the size, stage and prognosis of colorectal cancer, and may provide a new molecular marker for early diagnosis of colorectal cancer. Tiny RNA (mi RNA) can be used as a tumor suppressor or oncogene. Studies have shown that mi RNA-141 levels are up-regulated in colorectal cancer tissue samples. A number of possible target targets of mi RX14, including the tumor suppressor gene MAP2K4., were observed in the experiment. The purpose of this study was to investigate the role of mi Rc 141 in promoting the proliferation of colorectal cancer. Methods RT-PCR was used to detect the mRNA expression of mi-ru-141 and MAP2K4 in 20 cases of colorectal adenocarcinoma and their matched surrounding normal tissues. To analyze the correlation between mi rm 141 and TNM stage, to analyze the effect of inhibiting or increasing the expression of mi rm 141 on the proliferation of colorectal adenocarcinoma cell line by mi Rc 141 inhibitor or mi Rm 141 mimics, and to inhibit the expression of MAP2K4 by si RNA, and the expression of MAP2K4 in colorectal adenocarcinoma cell line was inhibited by si RNA. To investigate the role of MAP2K4 protein in the proliferation of colorectal cancer cells induced by mi Rc 141. Immunohistochemical staining was used to detect the expression of MAP2K4 protein and Western blot was used to detect the expression of MAP2K4 protein in colorectal cancer tissues and normal tissues. The proliferation of colorectal cancer cells was detected by MTT method. Results the expression level of miR-141 m RNA of miR-141 in TNM 鈪，

本文編號：2458544