宫颈癌中P21、Ki-67和IMP3基因表达的研究
发布时间：2019-05-16 23:36
【摘要】：研究目的:宫颈癌是最常见的妇科恶性肿瘤之一。原位癌高发年龄为30-35岁,浸润癌为45-55岁,近年来其发病有年轻化的趋势。因此宫颈癌的早期诊断、预后评估及各种治疗方法是人们研究的焦点。本研究拟检测P21、Ki-67和IMP3蛋白在CIN和宫颈癌组织及宫颈癌细胞株中的表达强弱以及是否有一致的表达关系。研究方法:(1)细胞培养与处理:从人正常宫颈组织中分离的宫颈上皮细胞(HCEC)及人宫颈癌细胞系的C33a、He La、Si Ha,于抗生素(链霉素和100U/ml青霉素)的RPMll640和含10%胎牛血清的培养基中培养,至细胞生长至80-90%的密度时,用TRIZOL及RIPA裂解液提取细胞裂解产物,行Real-time PCR技术检测P21、Ki-67和IMP3的m RNA表达水平以及Westernblot法检测P21、Ki-67和IMP3的蛋白水平。(2)免疫组化技术(IHC):免疫组织化学法检测30例正常宫颈组织、30例CIN、30例宫颈癌组织中p21、Ki-67和IMP3蛋白的表达情况。研究结果:He La、Si Ha、C33a细胞的P21、Ki-67和IMP3的m RNA及蛋白表达水平均高于人正常宫颈上皮细胞。正常宫颈组织中P21、Ki-67和IMP3的表达很弱,在CIN组织中P21、Ki-67和IMP3的表达阳性,而宫颈癌组织中P21、Ki-67和IMP3的表达呈强阳性。结论及意义:P21、Ki-67和IMP3蛋白在宫颈癌组织中有表达很强,在正常宫颈组织中却表达很弱,提示这三个基因可作为宫颈癌发生发展的共同检测指标,为宫颈癌的发生发展研究及靶向治疗提供一定的理论依据。
[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，
本文编号：2478640
[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，
本文编号：2478640
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