【摘要】：研究目的:宮頸癌是最常見的婦科惡性腫瘤之一。原位癌高發(fā)年齡為30-35歲,浸潤癌為45-55歲,近年來其發(fā)病有年輕化的趨勢。因此宮頸癌的早期診斷、預(yù)后評估及各種治療方法是人們研究的焦點(diǎn)。本研究擬檢測P21、Ki-67和IMP3蛋白在CIN和宮頸癌組織及宮頸癌細(xì)胞株中的表達(dá)強(qiáng)弱以及是否有一致的表達(dá)關(guān)系。研究方法:(1)細(xì)胞培養(yǎng)與處理:從人正常宮頸組織中分離的宮頸上皮細(xì)胞(HCEC)及人宮頸癌細(xì)胞系的C33a、He La、Si Ha,于抗生素(鏈霉素和100U/ml青霉素)的RPMll640和含10%胎牛血清的培養(yǎng)基中培養(yǎng),至細(xì)胞生長至80-90%的密度時(shí),用TRIZOL及RIPA裂解液提取細(xì)胞裂解產(chǎn)物,行Real-time PCR技術(shù)檢測P21、Ki-67和IMP3的m RNA表達(dá)水平以及Westernblot法檢測P21、Ki-67和IMP3的蛋白水平。(2)免疫組化技術(shù)(IHC):免疫組織化學(xué)法檢測30例正常宮頸組織、30例CIN、30例宮頸癌組織中p21、Ki-67和IMP3蛋白的表達(dá)情況。研究結(jié)果:He La、Si Ha、C33a細(xì)胞的P21、Ki-67和IMP3的m RNA及蛋白表達(dá)水平均高于人正常宮頸上皮細(xì)胞。正常宮頸組織中P21、Ki-67和IMP3的表達(dá)很弱,在CIN組織中P21、Ki-67和IMP3的表達(dá)陽性,而宮頸癌組織中P21、Ki-67和IMP3的表達(dá)呈強(qiáng)陽性。結(jié)論及意義:P21、Ki-67和IMP3蛋白在宮頸癌組織中有表達(dá)很強(qiáng),在正常宮頸組織中卻表達(dá)很弱,提示這三個(gè)基因可作為宮頸癌發(fā)生發(fā)展的共同檢測指標(biāo),為宮頸癌的發(fā)生發(fā)展研究及靶向治療提供一定的理論依據(jù)。
[Abstract]:Objective: cervical cancer is one of the most common gynecological malignant tumors. The high incidence age of carcinoma in situ was 30 years old, and that of invasive carcinoma was 45 years old. In recent years, the incidence of carcinoma in situ was younger than that of invasive carcinoma. Therefore, the early diagnosis, prognosis evaluation and various treatment methods of cervical cancer are the focus of people's research. The purpose of this study was to detect the expression of P21Ki67 and IMP3 proteins in CIN and cervical cancer tissues and cervical cancer cell lines and whether there was a consistent expression relationship. Methods: (1) Cell culture and treatment: cervical epithelial cells (HCEC) isolated from human normal cervical tissues and C33a, he La,Si Ha, of human cervical cancer cell lines The cells were cultured in RPMll640 of antibiotics (streptomycin and 100U/ml penicillin) and medium containing 10% fetal bovine serum. when the cells grew to a density of 80 鈮，

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