FISH技术检测宫颈癌组织中hTERC、HER-2基因表达及其临床意义
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摘要
目的应用FISH技术检测宫颈组织中hTERC、HER-2基因的表达,分析其与临床病理参数之间的关系,探讨hTERC、HER-2在宫颈癌发生、发展中的可能作用。
方法收集2008年3月至2009年3月于青岛大学医学院附属医院妇科就诊的162例患者的宫颈脱落细胞标本,其中正常及炎症者45例,CINⅠ者29例,CINⅡ-Ⅲ者38例,宫颈癌50例。收集同期手术切除或活检的宫颈组织标本63例,其中病理结果证实正常及炎症者13例,CINⅠ者12例,CINⅡ-Ⅲ者20例,宫颈癌者18例。采用五种制片方法,应用荧光原位杂交方法(FISH)检测所收集的细胞及组织中hTERC及HER-2基因的表达情况并比较五种制片方法之间的差异。
结果对于所检测的225例标本,其中直接涂片法12例、生理盐水法60例、低渗制片法20例、TCT制片法70例、石蜡包埋组织切片制片法63例,总杂交成功率为76.4%；TCT制片法、石蜡包埋组织切片制片法的杂交成功率与其他制片法的杂交成功率的差异有统计学意义(P<0.05),其中TCT制片法的杂交成功率最高,为87.1%.hTERC、HER-2基因在40例正常宫颈中不表达,而在其他病变组均有表达。hTERC基因在CINⅠ组、CINⅡ-Ⅲ组、宫颈癌组中的阳性表达率逐渐增高,差异具有统计学意义(P<0.05)；HER-2基因在CINⅠ组、CINⅡ-Ⅲ组、宫颈癌组中的阳性表达率分别为6.5%、28.6%、59.3%,依次逐渐增高,差异具有统计学意义(P<0.05).hTERC、HER-2基因的高表达与细胞分级均明显相关(P<0.05)。hTERC、HER-2基因在宫颈癌中的表达与肿瘤淋巴结的转移及分化程度均有明显相关(P<0.05),但与患者年龄、肿瘤大小、临床分期、组织学类型及病理类型无明显相关(P>0.05)。
结论对比五种FISH制片方法,在检测hTERC基因的宫颈癌筛查中,TCT制片法明显优于其他方法,而在指导宫颈病变及宫颈癌的治疗中,石蜡包埋组织切片法的实际意义更大。hTERC、HER-2基因高表达在子宫颈癌中发生发展中可能起重要作用,在子宫颈病变脱落细胞或组织中检测hTERC、HER-2基因可以作为子宫颈癌早期筛查、病情监测、评估预后的辅助指标。
Objective To explore the expression and their clinical significance of hTERC and HER-2 gene and to evaluate the effect and clinical significance of HER-2 and hTERC in cervical lesion.
Methods The fluorescent in situ hybridization (FISH) was used to detect the expression of the hTERC and HER-2 gene in cervical scraping samples and the Paraffin-embedded tissue sections with five Production Methods. The cervical scraping samples were divided into 45 in normal,29 in CIN I,38 in CINⅡ-Ⅲand 50 in cervical cancer. The Paraffin-embedded tissue sections were divided into 13 in normal,12 in CIN I,20 in CIN II-III and 18 in cervical cancer.
Results We detected 225 specimens, which including 12 cases of direct smear method,60 cases of saline method,20 cases of low osmotic method,70 cases of TCT production method and 63 cases of paraffin-embedded tissue sections method. The total hybridization success rate is 76.4%; The achievement ratio of the hybridization of hTERC and HER-2 gene for the direct smear method, the saline method, the TCT production method, the low osmotic method and the Paraffin-embedded tissue section method were 8.3%,65%,55%,87.1% and 85.7 %，respectively. The achievement ratio of TCT production method was higher than that of the other four group. The low osmotic method is better than others in the background of the cleanliness, the cell morphology and the number of the bare nuclei. The Paraffin-embedded tissue section method is the best in the satisfaction rate of the fluorescence signal. The hTERC and HER-2 gene in 40 cases of normal cervix was not expressed, while in other lesions were expressed. The hTERC gene in CIN I group, CINⅡ-Ⅲgroup, cervical cancer in the group positive expression rate are 16.1%,47.6% and 91.5% and the difference was statistically significant (P<0.05); The HER-2 gene in CIN I group, CIN II-III group, cervical cancer in the group positive expression rate are 6.5%,28.6% and 59.3% and the difference was statistically significant (P<0.05). The hTERC and HER-2 gene overexpression significantly associated with cell grade (P<0.05). And the hTERC and HER-2 gene in cervical carcinoma with tumor lymph node metastasis and differentiation of the degree of correlation (P<0.05). But the patient's age, tumor size, clinical stage, histological type and pathological types were non-significant correlation (P> 0.05).
Conclusion The TCT production method is better than other methods in the hTERC and HER-2 gene detecting of the cervical cancer screening. Otherwise, during the treatment guidance of the cervical lesions and the cervical cancer the Paraffin-embedded tissue section method is least at the damage of the chromosome and is the best at the practical significance. The hTERC and HER-2 gene were highly expressed in the occurrence and development of cervical cancer They may play an important role in cervical lesions detected in cervical carcinogenesis.The hTERC and HER-2 gene of cervical scraping samples could be used as a better tumor index for early cervical cancer screening and predicate the advancement and evaluate poor prognosis of cervical lesion.
方法收集2008年3月至2009年3月于青岛大学医学院附属医院妇科就诊的162例患者的宫颈脱落细胞标本,其中正常及炎症者45例,CINⅠ者29例,CINⅡ-Ⅲ者38例,宫颈癌50例。收集同期手术切除或活检的宫颈组织标本63例,其中病理结果证实正常及炎症者13例,CINⅠ者12例,CINⅡ-Ⅲ者20例,宫颈癌者18例。采用五种制片方法,应用荧光原位杂交方法(FISH)检测所收集的细胞及组织中hTERC及HER-2基因的表达情况并比较五种制片方法之间的差异。
结果对于所检测的225例标本,其中直接涂片法12例、生理盐水法60例、低渗制片法20例、TCT制片法70例、石蜡包埋组织切片制片法63例,总杂交成功率为76.4%；TCT制片法、石蜡包埋组织切片制片法的杂交成功率与其他制片法的杂交成功率的差异有统计学意义(P<0.05),其中TCT制片法的杂交成功率最高,为87.1%.hTERC、HER-2基因在40例正常宫颈中不表达,而在其他病变组均有表达。hTERC基因在CINⅠ组、CINⅡ-Ⅲ组、宫颈癌组中的阳性表达率逐渐增高,差异具有统计学意义(P<0.05)；HER-2基因在CINⅠ组、CINⅡ-Ⅲ组、宫颈癌组中的阳性表达率分别为6.5%、28.6%、59.3%,依次逐渐增高,差异具有统计学意义(P<0.05).hTERC、HER-2基因的高表达与细胞分级均明显相关(P<0.05)。hTERC、HER-2基因在宫颈癌中的表达与肿瘤淋巴结的转移及分化程度均有明显相关(P<0.05),但与患者年龄、肿瘤大小、临床分期、组织学类型及病理类型无明显相关(P>0.05)。
结论对比五种FISH制片方法,在检测hTERC基因的宫颈癌筛查中,TCT制片法明显优于其他方法,而在指导宫颈病变及宫颈癌的治疗中,石蜡包埋组织切片法的实际意义更大。hTERC、HER-2基因高表达在子宫颈癌中发生发展中可能起重要作用,在子宫颈病变脱落细胞或组织中检测hTERC、HER-2基因可以作为子宫颈癌早期筛查、病情监测、评估预后的辅助指标。
Objective To explore the expression and their clinical significance of hTERC and HER-2 gene and to evaluate the effect and clinical significance of HER-2 and hTERC in cervical lesion.
Methods The fluorescent in situ hybridization (FISH) was used to detect the expression of the hTERC and HER-2 gene in cervical scraping samples and the Paraffin-embedded tissue sections with five Production Methods. The cervical scraping samples were divided into 45 in normal,29 in CIN I,38 in CINⅡ-Ⅲand 50 in cervical cancer. The Paraffin-embedded tissue sections were divided into 13 in normal,12 in CIN I,20 in CIN II-III and 18 in cervical cancer.
Results We detected 225 specimens, which including 12 cases of direct smear method,60 cases of saline method,20 cases of low osmotic method,70 cases of TCT production method and 63 cases of paraffin-embedded tissue sections method. The total hybridization success rate is 76.4%; The achievement ratio of the hybridization of hTERC and HER-2 gene for the direct smear method, the saline method, the TCT production method, the low osmotic method and the Paraffin-embedded tissue section method were 8.3%,65%,55%,87.1% and 85.7 %，respectively. The achievement ratio of TCT production method was higher than that of the other four group. The low osmotic method is better than others in the background of the cleanliness, the cell morphology and the number of the bare nuclei. The Paraffin-embedded tissue section method is the best in the satisfaction rate of the fluorescence signal. The hTERC and HER-2 gene in 40 cases of normal cervix was not expressed, while in other lesions were expressed. The hTERC gene in CIN I group, CINⅡ-Ⅲgroup, cervical cancer in the group positive expression rate are 16.1%,47.6% and 91.5% and the difference was statistically significant (P<0.05); The HER-2 gene in CIN I group, CIN II-III group, cervical cancer in the group positive expression rate are 6.5%,28.6% and 59.3% and the difference was statistically significant (P<0.05). The hTERC and HER-2 gene overexpression significantly associated with cell grade (P<0.05). And the hTERC and HER-2 gene in cervical carcinoma with tumor lymph node metastasis and differentiation of the degree of correlation (P<0.05). But the patient's age, tumor size, clinical stage, histological type and pathological types were non-significant correlation (P> 0.05).
Conclusion The TCT production method is better than other methods in the hTERC and HER-2 gene detecting of the cervical cancer screening. Otherwise, during the treatment guidance of the cervical lesions and the cervical cancer the Paraffin-embedded tissue section method is least at the damage of the chromosome and is the best at the practical significance. The hTERC and HER-2 gene were highly expressed in the occurrence and development of cervical cancer They may play an important role in cervical lesions detected in cervical carcinogenesis.The hTERC and HER-2 gene of cervical scraping samples could be used as a better tumor index for early cervical cancer screening and predicate the advancement and evaluate poor prognosis of cervical lesion.
引文
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