hTERC、PCNA及HPV在宫颈病变中的表达及意义
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摘要
【目的】通过检测人端粒酶RNA (human telomerase RNA component,hTERC或hTR)和增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)在不同病变宫颈液基细胞学(Thinprep cytologic test,TCT)标本和组织中的表达,结合高危型人乳头瘤病毒(high risk human papillomaviruses,HR-HPV)检测结果,分析hTERC、PCNA及HR-HPV的相关性,讨论其与宫颈癌发生、发展的关系及临床意义,为宫颈癌的早期诊断和治疗提供新的实验理论依据。
【方法】分别收集宫颈上皮细胞内瘤变(cervical intraepithelia neoplasia, CIN)CINI、CINII、CINIII、浸润性宫颈癌(invasive cervical cancer, ICC)脱落细胞标本及相应的组织标本90例,其中CINI30例、CINII19例、CINIII 17例及ICC组织标本24例,以慢性宫颈炎症15例作为对照组。分别采用荧光原位杂交(fluorescence in situ hybridization ,FISH)检测hTERC基因在CINI、CINII、CINIII、浸润性宫颈癌(invasive cervical cancer, ICC)脱落细胞标本中的表达;采用免疫组织化学S-P(Streptavidin-peroxidase)法检测PCNA在CINI、CINII、CINIII、浸润性宫颈癌(invasive cervical cancer, ICC)组织标本中的表达情况;HR-HPV检测采用杂交捕获Ⅱ代(hybridization capture-Ⅱ,HC-Ⅱ)的方法,取上述病人的宫颈分泌物标本,检测高危型HPV(HR-HPV)病毒的载量。
【结果】1、hTERC基因在CINⅠ、Ⅱ、Ⅲ、ICC中的阳性表达率逐渐增高;与慢性宫颈炎症组相比,hTERC基因在CINⅡ、Ⅲ、ICC中的表达增高,差异具有显著性(P<0.01);CINⅠ、CINⅡ与ICC比较,hTERC的表达差异具有显著性(P<0.01);而CINⅢ与ICC比较,hTERC基因的表达无显著性差异(P >0.05)。
2、PCNA在CINⅡ、CINⅢ和ICC中的阳性表达率逐渐增高,与慢性宫颈炎症组比较,差异具有显著性(P<0.05);CINⅠ与慢性宫颈炎症组比较,无显著性差异(P >0.05)。CINⅢ和ICC与CINⅡ比较,PCNA表达的阳性率增高,有显著性差异(P<0.05);CINⅢ和ICC比较,PCNA表达的阳性率也存在显著性差异(P<0.05)。在早期浸润癌中,Ⅰa-Ⅰb和Ⅱa比较,PCNA表达的阳性率也存在显著性差异(P<0.05)。
3、HR-HPV在宫颈CINⅠ、CINⅡ、CINⅢ、ICC宫颈分泌物中的阳性表达率逐渐增高,与慢性宫颈炎症组比较,差异具有显著性(P<0.01);与CINⅠ比较,CINⅡ、CINⅢ与ICC HR-HPV的阳性表达率增强,存在显著性差异(P<0.01)。CINⅢ与ICC比较,其阳性表达率增强,但无显著性差异(P >0.05)。
4、hTERC基因和PCNA蛋白及HR-HPV在宫颈病变中的表达随病变程度的增高具有逐步上升的趋势。采用Bivariate双变量相关分析,得出hTERC和PCNA及HR-HPV之间的相关系数分别为0.903和0.922(P<0.05);PCNA及HR-HPV之间的相关系数rs为0.943(P<0.05),三者之间存在高度相关。
【结论】hTERC、PCNA及HR-HPV在CIN和ICC中显著高表达,可能为宫颈癌发展过程中重要的分子事件。hTERC和PCNA及HR-HPV联合检测可能为宫颈癌的早期诊断和治疗提供临床依据。
Objective: Through investigation of the expression of hTERC and PCNA in the sample of cervical TCT and tissue of different cervical diseases, binding the expression of HR-HPV, to analyze the correlation among hTERC and PCNA and HR-HPV. According to their relation, the occurrence and development of cervical carcinoma and clinical significance are discussed. The findings provided new experiment theoretical evidence for early diagnosis and therapy of cervical carcinoma.
Method:To collect 105 cases of cervical TCT samples and their tissue samples,chronic inflammation 15 cases,CINⅠ30 CINⅡ19 cases,CINⅢ17 cases,and early invasive cervical cancer 24 cases. We measured:1) hTERC in TCT sample by FISH method; and PCNA in tissue samples by Immunohistochemical method; 3)the load quantity of HR-HPV in secretion of cervical by HC-Ⅱ.
Result: 1. The significant statistic difference was found in the expression of hTERC in CINⅠ、Ⅱ、Ⅲand CC after comparing with the group of chronic cervicitis(P<0.01). Comparing with the group of CINⅠ, the expression of hTERC in CINⅡ、Ⅲand CC also found to have significant difference(P<0.01).While difference between the group of CINⅢand CC are not significant(P >0.05).
2. Comparing with the group of the chronic cervicitis, the expression of PCNA in CINⅡ、Ⅲand CC had significant difference(P<0.05); There wasn’t significant difference between the group of CINⅠand the group of the chronic cervicitis (P>0.05). The expression of PCNA in CINⅢand CC was significantly different from the expression of PCNA in CINⅡ(P<0.05). The significant difference in expression of PCNA also exists between the group of CINⅢand CC(P <0.05). The expression of PCNA inⅠa-Ⅰb andⅡa of early ICC was significantly differenence(P<0.05).
3. Comparing with the group of the chronic cervicitis, the expression of HR-HPV in CIN groups and CC had significant difference(P<0.05); Comparing with CINⅠ, significance difference exists in the expression of CINⅡ, CINⅢand CC (P<0.05) . Compared with CC, the positive expression of HR-HPV of CINⅢis increasing, but the difference wasn’t significant (P>0.05).
4. As the developing of the of cervical carcinoma ,the positive rates of hTERC and PCNA and HR-HPV in the group of chronic cervicities CIN and CC presented a trend of gradual rising. Through the way of Bivarite,a closely positive correlation was found among hTERC and PCNA and HR-HPV(rs=0.903, P<0.05 hTERC vs PCNA P<0.05 ; rs= 0.922 ,hTERC vs HR-HPV, P<0.05 ;rs=0.943, P<0.05, PCNA vs HR-HPV)
Conclusion: High expression of hTERC and PCNA and HR-HPV in CIN and CC,may be important molecule signs of cervical carcinoma. The combined detection made by hTERC, PCNA and HR-HPV may offer clinical evidences in early diagnosis and therapy in CC.
【方法】分别收集宫颈上皮细胞内瘤变(cervical intraepithelia neoplasia, CIN)CINI、CINII、CINIII、浸润性宫颈癌(invasive cervical cancer, ICC)脱落细胞标本及相应的组织标本90例,其中CINI30例、CINII19例、CINIII 17例及ICC组织标本24例,以慢性宫颈炎症15例作为对照组。分别采用荧光原位杂交(fluorescence in situ hybridization ,FISH)检测hTERC基因在CINI、CINII、CINIII、浸润性宫颈癌(invasive cervical cancer, ICC)脱落细胞标本中的表达;采用免疫组织化学S-P(Streptavidin-peroxidase)法检测PCNA在CINI、CINII、CINIII、浸润性宫颈癌(invasive cervical cancer, ICC)组织标本中的表达情况;HR-HPV检测采用杂交捕获Ⅱ代(hybridization capture-Ⅱ,HC-Ⅱ)的方法,取上述病人的宫颈分泌物标本,检测高危型HPV(HR-HPV)病毒的载量。
【结果】1、hTERC基因在CINⅠ、Ⅱ、Ⅲ、ICC中的阳性表达率逐渐增高;与慢性宫颈炎症组相比,hTERC基因在CINⅡ、Ⅲ、ICC中的表达增高,差异具有显著性(P<0.01);CINⅠ、CINⅡ与ICC比较,hTERC的表达差异具有显著性(P<0.01);而CINⅢ与ICC比较,hTERC基因的表达无显著性差异(P >0.05)。
2、PCNA在CINⅡ、CINⅢ和ICC中的阳性表达率逐渐增高,与慢性宫颈炎症组比较,差异具有显著性(P<0.05);CINⅠ与慢性宫颈炎症组比较,无显著性差异(P >0.05)。CINⅢ和ICC与CINⅡ比较,PCNA表达的阳性率增高,有显著性差异(P<0.05);CINⅢ和ICC比较,PCNA表达的阳性率也存在显著性差异(P<0.05)。在早期浸润癌中,Ⅰa-Ⅰb和Ⅱa比较,PCNA表达的阳性率也存在显著性差异(P<0.05)。
3、HR-HPV在宫颈CINⅠ、CINⅡ、CINⅢ、ICC宫颈分泌物中的阳性表达率逐渐增高,与慢性宫颈炎症组比较,差异具有显著性(P<0.01);与CINⅠ比较,CINⅡ、CINⅢ与ICC HR-HPV的阳性表达率增强,存在显著性差异(P<0.01)。CINⅢ与ICC比较,其阳性表达率增强,但无显著性差异(P >0.05)。
4、hTERC基因和PCNA蛋白及HR-HPV在宫颈病变中的表达随病变程度的增高具有逐步上升的趋势。采用Bivariate双变量相关分析,得出hTERC和PCNA及HR-HPV之间的相关系数分别为0.903和0.922(P<0.05);PCNA及HR-HPV之间的相关系数rs为0.943(P<0.05),三者之间存在高度相关。
【结论】hTERC、PCNA及HR-HPV在CIN和ICC中显著高表达,可能为宫颈癌发展过程中重要的分子事件。hTERC和PCNA及HR-HPV联合检测可能为宫颈癌的早期诊断和治疗提供临床依据。
Objective: Through investigation of the expression of hTERC and PCNA in the sample of cervical TCT and tissue of different cervical diseases, binding the expression of HR-HPV, to analyze the correlation among hTERC and PCNA and HR-HPV. According to their relation, the occurrence and development of cervical carcinoma and clinical significance are discussed. The findings provided new experiment theoretical evidence for early diagnosis and therapy of cervical carcinoma.
Method:To collect 105 cases of cervical TCT samples and their tissue samples,chronic inflammation 15 cases,CINⅠ30 CINⅡ19 cases,CINⅢ17 cases,and early invasive cervical cancer 24 cases. We measured:1) hTERC in TCT sample by FISH method; and PCNA in tissue samples by Immunohistochemical method; 3)the load quantity of HR-HPV in secretion of cervical by HC-Ⅱ.
Result: 1. The significant statistic difference was found in the expression of hTERC in CINⅠ、Ⅱ、Ⅲand CC after comparing with the group of chronic cervicitis(P<0.01). Comparing with the group of CINⅠ, the expression of hTERC in CINⅡ、Ⅲand CC also found to have significant difference(P<0.01).While difference between the group of CINⅢand CC are not significant(P >0.05).
2. Comparing with the group of the chronic cervicitis, the expression of PCNA in CINⅡ、Ⅲand CC had significant difference(P<0.05); There wasn’t significant difference between the group of CINⅠand the group of the chronic cervicitis (P>0.05). The expression of PCNA in CINⅢand CC was significantly different from the expression of PCNA in CINⅡ(P<0.05). The significant difference in expression of PCNA also exists between the group of CINⅢand CC(P <0.05). The expression of PCNA inⅠa-Ⅰb andⅡa of early ICC was significantly differenence(P<0.05).
3. Comparing with the group of the chronic cervicitis, the expression of HR-HPV in CIN groups and CC had significant difference(P<0.05); Comparing with CINⅠ, significance difference exists in the expression of CINⅡ, CINⅢand CC (P<0.05) . Compared with CC, the positive expression of HR-HPV of CINⅢis increasing, but the difference wasn’t significant (P>0.05).
4. As the developing of the of cervical carcinoma ,the positive rates of hTERC and PCNA and HR-HPV in the group of chronic cervicities CIN and CC presented a trend of gradual rising. Through the way of Bivarite,a closely positive correlation was found among hTERC and PCNA and HR-HPV(rs=0.903, P<0.05 hTERC vs PCNA P<0.05 ; rs= 0.922 ,hTERC vs HR-HPV, P<0.05 ;rs=0.943, P<0.05, PCNA vs HR-HPV)
Conclusion: High expression of hTERC and PCNA and HR-HPV in CIN and CC,may be important molecule signs of cervical carcinoma. The combined detection made by hTERC, PCNA and HR-HPV may offer clinical evidences in early diagnosis and therapy in CC.
引文
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