宫颈病变中P16～（INK4a）、CyclinE、CDK2的表达以及与HPV_（16）感染关系的研究
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摘要
目的 探讨 P16INK4a、CyclinE、CDK2 在各类宫颈病变中的表达以及与
HPV16 感染关系的研究，为临床宫颈癌及癌前病变的诊断及筛查提供理论依
据及寻找一种经济有效、快速的筛查方法。
方法 应用组织微阵列方法和免疫组化法对 15 例正常宫颈、10 例宫颈
湿疣、45 例宫颈上皮内瘤样病变、20 例宫颈浸润癌行 P16INK4a、CyclinE、
CDK2 的测定（SP 法检测 P16INK4a、CyclinE；SABC 检测 CDK2），用组织微阵
列方法及原位杂交行 HPV16CDNA 的测定。
结果 宫颈癌组的产次显著高于其它各组宫颈病变组（P<0.05）。随病
变进展 HPV16CDNA 表达增加，与宫颈癌及宫颈上皮内瘤变呈正相关
（P<0.001）。P16INK4a、CyclinE、CDK2 在正常宫颈中无表达，而随着宫颈病
变的进展，P16INK4a、CyclinE、CDK2 的表达率显著升高（P<0.001）, 且与
HPV16有相关性（相关系数分别为 0.363、0.683、0.338）。三者在宫颈癌中
的表达率分别为 95%、60%、85%。
结论 组织微阵列技术适用于大样本量的检测，是一种高效、快速、
可比性强的分子生物学研究方法。宫颈癌与多产有关。P16INK4a、CyclinE、
CDK2 在一定程度上可反映 HPV 的感染程度，可作为宫颈癌筛查的指标。灵
敏度大小依次为 P16INK4a 、CDK2、CyclinE。特异度最高的是 CyclinE。P16INK4a、
CyclinE、CDK2 在宫颈癌的发生发展中可能起着重要的作用。
Objective To study the expression and significance of P16INK4a、
CyclinE、CDK2 in cervical lesions and correlation between HPV16. In order
to offer an ecomomic valid rapid method which can be used for clinical
diagnosis and screening of precancerous and earlier carcinomas.
Method In immunohistochemistry detection P16INK4A , CyclinE,
CDK2 of expression（Detection P16INK4A ,CyclinE using SP method;
Detection CDK2 using SABC method）and in situ hybridization detection
HPV16 in 15 nomal cervix,10 cervical condyloma ,45 CINs and 20 cervical
carcinoma Using Tissue Microarra.
Result The para of cervical cancer was significantly higher than those
of other cervical lesions（P<0.05）.The expresion of HPV16CDNA increases
following the development of cervical lesions. The expresion of HPV16CDNA
and cervical carcinoma 、CINs has positive correlation（P<0.001）. We found no
cases with expression of P16INK4a、CyclinE、CDK2 in nomal cervix.The
expression rates of P16INK4a、CyclinE、CDK2 was significantly higher following
the development of cervical lesions（P<0.001）. Close positive correlation was
observed between the expression of P16INK4a 、 CyclinE 、 CDK2 and
HPV16CDNA(rs were 0.363、0.683、0.338). The expression rates of P16INK4a、
CyclinE、CDK2 were 95%,60%, and 85% in cervical cancer。
Conclusion Tissue microarray is an efficient technique of molecular
II
biology.Cervical cancer is an disease correlationing between more para. The
expression of P16INK4a、CyclinE、CDK2 may reflect the seriousness of
HPV infection .They can become a screening test. The overexpression of
P16INK4a、CyclinE、CDK2 may play an important role in cervical cancer.
HPV16 感染关系的研究，为临床宫颈癌及癌前病变的诊断及筛查提供理论依
据及寻找一种经济有效、快速的筛查方法。
方法 应用组织微阵列方法和免疫组化法对 15 例正常宫颈、10 例宫颈
湿疣、45 例宫颈上皮内瘤样病变、20 例宫颈浸润癌行 P16INK4a、CyclinE、
CDK2 的测定（SP 法检测 P16INK4a、CyclinE；SABC 检测 CDK2），用组织微阵
列方法及原位杂交行 HPV16CDNA 的测定。
结果 宫颈癌组的产次显著高于其它各组宫颈病变组（P<0.05）。随病
变进展 HPV16CDNA 表达增加，与宫颈癌及宫颈上皮内瘤变呈正相关
（P<0.001）。P16INK4a、CyclinE、CDK2 在正常宫颈中无表达，而随着宫颈病
变的进展，P16INK4a、CyclinE、CDK2 的表达率显著升高（P<0.001）, 且与
HPV16有相关性（相关系数分别为 0.363、0.683、0.338）。三者在宫颈癌中
的表达率分别为 95%、60%、85%。
结论 组织微阵列技术适用于大样本量的检测，是一种高效、快速、
可比性强的分子生物学研究方法。宫颈癌与多产有关。P16INK4a、CyclinE、
CDK2 在一定程度上可反映 HPV 的感染程度，可作为宫颈癌筛查的指标。灵
敏度大小依次为 P16INK4a 、CDK2、CyclinE。特异度最高的是 CyclinE。P16INK4a、
CyclinE、CDK2 在宫颈癌的发生发展中可能起着重要的作用。
Objective To study the expression and significance of P16INK4a、
CyclinE、CDK2 in cervical lesions and correlation between HPV16. In order
to offer an ecomomic valid rapid method which can be used for clinical
diagnosis and screening of precancerous and earlier carcinomas.
Method In immunohistochemistry detection P16INK4A , CyclinE,
CDK2 of expression（Detection P16INK4A ,CyclinE using SP method;
Detection CDK2 using SABC method）and in situ hybridization detection
HPV16 in 15 nomal cervix,10 cervical condyloma ,45 CINs and 20 cervical
carcinoma Using Tissue Microarra.
Result The para of cervical cancer was significantly higher than those
of other cervical lesions（P<0.05）.The expresion of HPV16CDNA increases
following the development of cervical lesions. The expresion of HPV16CDNA
and cervical carcinoma 、CINs has positive correlation（P<0.001）. We found no
cases with expression of P16INK4a、CyclinE、CDK2 in nomal cervix.The
expression rates of P16INK4a、CyclinE、CDK2 was significantly higher following
the development of cervical lesions（P<0.001）. Close positive correlation was
observed between the expression of P16INK4a 、 CyclinE 、 CDK2 and
HPV16CDNA(rs were 0.363、0.683、0.338). The expression rates of P16INK4a、
CyclinE、CDK2 were 95%,60%, and 85% in cervical cancer。
Conclusion Tissue microarray is an efficient technique of molecular
II
biology.Cervical cancer is an disease correlationing between more para. The
expression of P16INK4a、CyclinE、CDK2 may reflect the seriousness of
HPV infection .They can become a screening test. The overexpression of
P16INK4a、CyclinE、CDK2 may play an important role in cervical cancer.
引文
[1] 郎景和. CIN 的诊断和治疗.中华妇产科杂志,2001，36(5):261
[2] Food Nutrition And The Prevention of Cancer:a global perspective. First
published 1997 byAICR，301-308
[3] Anttila A,Pukkala E,Soderman,et al.Effect of organized screening on
cervical cancer incidence and mortality in Finland,1963-1995:Recent incease in
cervicalcancer incidence.Int J Cancer,1999,83:59-65
[4] 高进.肿瘤学基础与研究方法.北京：人民卫生出版社，1999，
[5] 李连第,鲁风珠,张思维,等。中国恶性肿瘤死亡率 20 年变化趋势和年近
期预测分析，中华肿瘤杂志，1997，19，3-9
[6] Duensing S, Munger K. Centrosome abnormalities and genomic instability
induced by human papillomavirus oncoproteins. Prog Cell Cycle Res, 2003,5:383-91
[7] 戎寿地，陈汶，吴令英，等.山西省襄垣县宫颈癌危险因素分析.中华
预防医学杂志，2002，36（1）：41-43
[8] Kaznelson DW, Bruun S, Monrad A, et al. Simultaneous human
papilloma virus type 16 E7 and cdk inhibitor p21 expression induces apoptosis and
cathepsin B activation. Virology, 2004 320(2):301-12.
[9] 石一复等.子宫颈疾病.北京:人民卫生出版社,1999,58
[10] 刘树范.浅析巴氏五类法与TBS 描述性诊断报告方式.中国实用妇科与
产科杂志,2003，19 (3):135-137
[11] 唐怀好,王毅,满红玲等.周期素依赖激酶 2 在非小细胞肺癌中的表达及
其临床意义.中国胸心血管外科临床杂志,2003,10(2):95
[12] 王伯沅，李玉松，黄高晟等主编《病理学技术》 人民卫生出版社，
2000
[13] Kononen J,Bubendorf L,Kallioniemi A,et al.Tissue microarrays for
high-throughput molecular profiling of tumor specimens.Nat Med,1998,4:844-847
[14] Bubendorf L,Kononen J,Koivisto P,et al.Survey of gene amplifications
during prostate cancer progression by high-throughput fluorescence in situ
hybridization on tissue microarrays.Cancer Res,1999,59:803-806
18
山西医科大学硕士学位论文
[15] Hausen ZH.Papillomaviruses in human cancer. Cancer,1987,59:1692-
1696
[16]Klaes R, Friedrich T, Spitkovsky D, et al. Overexpression of p16(INK4A)
as a specific marker for dysplastic and neoplastic epithelial cells of the cervix uteri.
Int J Cancer, 2001, 92: 276–84
[17] Kamb A. Cell-cycle regulators and cancer. Trends Genet, 1995, 11:136–40
[18] Caldas C, Hahn SA, da Costa LT, et al. Frequent somatic mutations and
homozygous deletions of the p16 (MTS1) gene in pancreatic adenocarcinoma. Nat
Genet, 1994, 8:27–32
[19] Parry D, Bates S, Mann DJ, et al. Lack of cyclin D-Cdk complexes in
Rb-negative cells correlates with high levels of p16INK4/MTS1 tumour suppressor
gene product. EMBO J, 1995, 14:503–11
[20] Agoff, S, Nicholas M.D. Lin, Patricia M.P.H., et al. p16INK4a Expression
Correlates with Degree of Cervical Neoplasia: A Comparison with Ki-67 Expression
and Detection of High-Risk HPV Types ,The United States and Canadian Academy
of Pathology, Inc. 2003,16(7):665-673
[21] Khleif SN, DeGregori J, Yee CL, et al. Inhibition of cyclin D-CDK4/
CDK6 activity is associated with an E2F-mediated induction of cyclin kinase
inhibitor activity. Proc NatAcad Sciences USA, 1996,93:4350–4
[22] Sahebali S, Depuydt CE, Segers K, et al. P16INK4a as an adjunct marker
in liquid-based cervical cytology. Int J Cancer， 2004，108(6):871-6
[23] Keyomarsi K,Oleary N, Molnar G et al. CyclinE, a potential prognostic
markerfor breast cancer.Cancer Res,1994,54:380-385
[24] Weaver EJ,Kovtich AJ,Bibbo M. CyclinE expression and early cervical
neoplasia in ThinPrep specimens.A feasibility study.Acta Cytol,2000,44(3):
301-304
[25] LilischkisR,BarczakE,MarwedelM,et al.Identification of new substrates of
cyclin-activated kinases in breast cancers.VerhDtschGesPathol,2001,85:269-274
[26] 钱德英.阴道镜在宫颈癌诊断中的应用.中国实用妇科与产科杂
志,2003，19(3):137-138
19
山西医科大学硕士学位论文
[27] Meisels A.Fortin R.Roy M.condylomatous lesions of the cervix,
Ⅱ.cytologic ,colposcopic and histopathlolgic study.Acta Cytol, 1997, 21:379-390
[28] Ricard RM.Amodified terminology for cervical interaepithial neoplasia.
Obstet Gynecol,1990,75:131-133
[2] Food Nutrition And The Prevention of Cancer:a global perspective. First
published 1997 byAICR，301-308
[3] Anttila A,Pukkala E,Soderman,et al.Effect of organized screening on
cervical cancer incidence and mortality in Finland,1963-1995:Recent incease in
cervicalcancer incidence.Int J Cancer,1999,83:59-65
[4] 高进.肿瘤学基础与研究方法.北京：人民卫生出版社，1999，
[5] 李连第,鲁风珠,张思维,等。中国恶性肿瘤死亡率 20 年变化趋势和年近
期预测分析，中华肿瘤杂志，1997，19，3-9
[6] Duensing S, Munger K. Centrosome abnormalities and genomic instability
induced by human papillomavirus oncoproteins. Prog Cell Cycle Res, 2003,5:383-91
[7] 戎寿地，陈汶，吴令英，等.山西省襄垣县宫颈癌危险因素分析.中华
预防医学杂志，2002，36（1）：41-43
[8] Kaznelson DW, Bruun S, Monrad A, et al. Simultaneous human
papilloma virus type 16 E7 and cdk inhibitor p21 expression induces apoptosis and
cathepsin B activation. Virology, 2004 320(2):301-12.
[9] 石一复等.子宫颈疾病.北京:人民卫生出版社,1999,58
[10] 刘树范.浅析巴氏五类法与TBS 描述性诊断报告方式.中国实用妇科与
产科杂志,2003，19 (3):135-137
[11] 唐怀好,王毅,满红玲等.周期素依赖激酶 2 在非小细胞肺癌中的表达及
其临床意义.中国胸心血管外科临床杂志,2003,10(2):95
[12] 王伯沅，李玉松，黄高晟等主编《病理学技术》 人民卫生出版社，
2000
[13] Kononen J,Bubendorf L,Kallioniemi A,et al.Tissue microarrays for
high-throughput molecular profiling of tumor specimens.Nat Med,1998,4:844-847
[14] Bubendorf L,Kononen J,Koivisto P,et al.Survey of gene amplifications
during prostate cancer progression by high-throughput fluorescence in situ
hybridization on tissue microarrays.Cancer Res,1999,59:803-806
18
山西医科大学硕士学位论文
[15] Hausen ZH.Papillomaviruses in human cancer. Cancer,1987,59:1692-
1696
[16]Klaes R, Friedrich T, Spitkovsky D, et al. Overexpression of p16(INK4A)
as a specific marker for dysplastic and neoplastic epithelial cells of the cervix uteri.
Int J Cancer, 2001, 92: 276–84
[17] Kamb A. Cell-cycle regulators and cancer. Trends Genet, 1995, 11:136–40
[18] Caldas C, Hahn SA, da Costa LT, et al. Frequent somatic mutations and
homozygous deletions of the p16 (MTS1) gene in pancreatic adenocarcinoma. Nat
Genet, 1994, 8:27–32
[19] Parry D, Bates S, Mann DJ, et al. Lack of cyclin D-Cdk complexes in
Rb-negative cells correlates with high levels of p16INK4/MTS1 tumour suppressor
gene product. EMBO J, 1995, 14:503–11
[20] Agoff, S, Nicholas M.D. Lin, Patricia M.P.H., et al. p16INK4a Expression
Correlates with Degree of Cervical Neoplasia: A Comparison with Ki-67 Expression
and Detection of High-Risk HPV Types ,The United States and Canadian Academy
of Pathology, Inc. 2003,16(7):665-673
[21] Khleif SN, DeGregori J, Yee CL, et al. Inhibition of cyclin D-CDK4/
CDK6 activity is associated with an E2F-mediated induction of cyclin kinase
inhibitor activity. Proc NatAcad Sciences USA, 1996,93:4350–4
[22] Sahebali S, Depuydt CE, Segers K, et al. P16INK4a as an adjunct marker
in liquid-based cervical cytology. Int J Cancer， 2004，108(6):871-6
[23] Keyomarsi K,Oleary N, Molnar G et al. CyclinE, a potential prognostic
markerfor breast cancer.Cancer Res,1994,54:380-385
[24] Weaver EJ,Kovtich AJ,Bibbo M. CyclinE expression and early cervical
neoplasia in ThinPrep specimens.A feasibility study.Acta Cytol,2000,44(3):
301-304
[25] LilischkisR,BarczakE,MarwedelM,et al.Identification of new substrates of
cyclin-activated kinases in breast cancers.VerhDtschGesPathol,2001,85:269-274
[26] 钱德英.阴道镜在宫颈癌诊断中的应用.中国实用妇科与产科杂
志,2003，19(3):137-138
19
山西医科大学硕士学位论文
[27] Meisels A.Fortin R.Roy M.condylomatous lesions of the cervix,
Ⅱ.cytologic ,colposcopic and histopathlolgic study.Acta Cytol, 1997, 21:379-390
[28] Ricard RM.Amodified terminology for cervical interaepithial neoplasia.
Obstet Gynecol,1990,75:131-133