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目的:探讨宫颈癌中表皮生长因子受体(EGFR)、蛋白激酶B(PKB/Akt)的表达及EGFR胞外区基因突变的规律和临床意义,为宫颈癌的发生发展机制和靶向个性化治疗提供实验依据。方法:采用免疫组化SP法检测EGFR、Akt蛋白在16例正常宫颈组织、40例宫颈上皮内瘤变(CIN)组织和60例宫颈癌组织中的表达,并分析其与宫颈癌临床病理的关系。采用酚氯仿法抽提60例宫颈癌组织中DNA,PCR+DNA双向测序法检测EGFR胞外IV亚区基因突变。结果:正常宫颈组、CIN组及宫颈癌组中EGFR蛋白的阳性表达率分别为13.33%、43.33%及76.67%,Akt蛋白的阳性表达率分别为6.25%、35.00%及68.33%,差异有统计学意义(P<0.05)。宫颈癌组织中,EGFR表达水平与病理类型、细胞分化程度和临床分期有关(P<0.05),但与患者年龄和淋巴结转移无关(P>0.05);Akt表达水平与细胞分化程度和临床分期有关(P<0.05),但与患者年龄、病理类型和淋巴结转移无关(P>0.05)。60例宫颈癌中EGFR基因外显子15突变率为3.33%(2/60),突变类型G→A,未发现外显子14突变。宫颈癌组织中EGFR、Akt表达呈正相关性(r=0.556,P<0.05)。结论:EGFR、Akt可作为检测宫颈癌及癌前病变的重要参考指标,也为宫颈癌的多靶点联合治疗提供新思路;宫颈癌中EGFR胞外区可有基因突变,但突变率很低,关于其突变的规律和意义有待进一步研究。
Objective: To investigate the regularity and clinical significance of the expression of epidermal growth factor receptor (EGFR), protein kinase B (PKB / Akt) and gene mutations in extracellular region of cervical cancer, and to study the mechanism of cervical carcinogenesis and personalized targeting Treatment provides experimental evidence. Methods: Immunohistochemical SP method was used to detect the expression of EGFR and Akt protein in 16 cases of normal cervical tissue, 40 cases of cervical intraepithelial neoplasia (CIN) and 60 cases of cervical cancer, and analyzed its relationship with clinicopathological features relationship. DNA was extracted from 60 cases of cervical cancer tissues by phenol-chloroform method, and the gene mutations of extracellular IV subunit of EGFR were detected by PCR + DNA bi-directional sequencing. Results: The positive rates of EGFR protein in normal cervix, CIN group and cervical cancer group were 13.33%, 43.33% and 76.67% respectively, and the positive rates of Akt protein were 6.25%, 35.00% and 68.33%, respectively Significance (P <0.05). The expression of EGFR in cervical cancer was correlated with the pathological type, cell differentiation and clinical stage (P <0.05), but not with age and lymph node metastasis (P> 0.05). The expression level of Akt was correlated with cell differentiation and clinical stage (P <0.05), but not with age, pathological type and lymph node metastasis (P> 0.05). The mutation rate of exon 15 in EGFR gene was 3.33% (2/60) in 60 cases of cervical cancer, and the mutation type G → A did not find exon 14 mutation. The expression of EGFR and Akt in cervical cancer tissues was positively correlated (r = 0.556, P <0.05). Conclusion: EGFR and Akt can be used as important reference markers for detecting cervical cancer and precancerous lesions, and also provide new ideas for multi-target combination therapy of cervical cancer. The mutation of EGFR and Akt may be found in the extracellular region of cervical cancer, but the mutation rate is very low , The rules and significance of its mutation needs further study.