宫颈癌介入化疗前后核形态、AgNOR及PCNA的定量研究
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摘要
目的 探讨介入化疗对宫颈鳞癌细胞核形态、核仁组成区嗜银蛋白（AgNOR）、增殖细胞核抗原（PCNA）的影响。
方法 用免疫组化、HE 和嗜银染色对30例进行介入化疗前后的宫颈癌组织染色，并用真彩色图像分析系统测量肿瘤细胞的核面积、核周长、最大直径、AgNOR颗粒截面数、颗粒直径、颗粒面积、颗粒面积∕核面积以及PCNA阳性率。
结果 介入后宫颈癌细胞的核面积、核周长、核最大直径（分别为61.94um2、29.35 um、11.00 um）较介入前（分别为82.83um2、36.15um、17.8um）缩小，P＜0.05，有显著差异；通过计算得到的介入前核体积密度、核数密度、表面积密度分别为0.037、0.005、0.037，介入后的上述值分别为0.021、0.029、0.013，两者比较P＜0.05，有显著差异；通过计算得到的介入前核平均体积、核平均表面积、三维形状因子分别为9.69、4.6、1.094，介入后的上述值分别为4.120、0.961、0.713，两者比较P＜0.05，有显著差异。介入后AgNOR颗粒截面数、颗粒面积、颗粒直径、颗粒面积／核面积（分别为3.80个、3.26um2、1.08um、0.12）较介入前（分别为4.97个、7.98um2、0.72um、0.23）减少或缩小，P＜0.05，有显著差异。介入后PCNA积分为5.03较介入前（为3.23）减少，P＜0.05，有显著差异。
结论 1、介入化疗可以缩小宫颈癌细胞的核面积、核周长，并且介入化疗前后宫颈癌的组织结构参数（Vv、Sv、Nv、V、S、Rsv）存在显著差异。2、介入化疗可以减少AgNOR颗粒截面数和降低PCNA阳性率，表明介入化疗可以抑制宫颈癌细胞的增殖。
Objective: To study the effect of intra-arterial infusion chemotherapy on nuclear morphometry ,AgNOR and PCNA in cervical cancer .
Methods : 30 cervical cancer were examined for multiple biomarkers (nuclear morphometry ,AgNOR and PCNA) with quantitative compute processed imaging analysis before intra-arterial infusion chemotherapy and after intra-arterial infusion chemotherapy.
Results : Statistical analysis showed that there were significant differences between the groups (P <0.05). ⑴ There was significant differences in 9 parameters (Prmtr, At ,Dmax, Vv ,Nv , Sv , V, S, Rsv) between before and after intra-arterial infusion chemotherapy of nuclear morphometry . ⑵AgNOR counts of before and after intra-arterial infusion chemotherapy were 6.70± 0.92 and 4.05±0.62. After treatment ,the AgNOR configuration changed greatly . ⑶Positive rates of PCNA before and after intra-arterial infusion chemotherapy were significant difference .
Conclusion : Intera-arterial infusion chemotherapy can inhibit the proliferation of cervical cancer cell . It is recommended that intra-arterial infusion chemotherapy be employed to be one of combined treatments for cervical cancer.
方法 用免疫组化、HE 和嗜银染色对30例进行介入化疗前后的宫颈癌组织染色，并用真彩色图像分析系统测量肿瘤细胞的核面积、核周长、最大直径、AgNOR颗粒截面数、颗粒直径、颗粒面积、颗粒面积∕核面积以及PCNA阳性率。
结果 介入后宫颈癌细胞的核面积、核周长、核最大直径（分别为61.94um2、29.35 um、11.00 um）较介入前（分别为82.83um2、36.15um、17.8um）缩小，P＜0.05，有显著差异；通过计算得到的介入前核体积密度、核数密度、表面积密度分别为0.037、0.005、0.037，介入后的上述值分别为0.021、0.029、0.013，两者比较P＜0.05，有显著差异；通过计算得到的介入前核平均体积、核平均表面积、三维形状因子分别为9.69、4.6、1.094，介入后的上述值分别为4.120、0.961、0.713，两者比较P＜0.05，有显著差异。介入后AgNOR颗粒截面数、颗粒面积、颗粒直径、颗粒面积／核面积（分别为3.80个、3.26um2、1.08um、0.12）较介入前（分别为4.97个、7.98um2、0.72um、0.23）减少或缩小，P＜0.05，有显著差异。介入后PCNA积分为5.03较介入前（为3.23）减少，P＜0.05，有显著差异。
结论 1、介入化疗可以缩小宫颈癌细胞的核面积、核周长，并且介入化疗前后宫颈癌的组织结构参数（Vv、Sv、Nv、V、S、Rsv）存在显著差异。2、介入化疗可以减少AgNOR颗粒截面数和降低PCNA阳性率，表明介入化疗可以抑制宫颈癌细胞的增殖。
Objective: To study the effect of intra-arterial infusion chemotherapy on nuclear morphometry ,AgNOR and PCNA in cervical cancer .
Methods : 30 cervical cancer were examined for multiple biomarkers (nuclear morphometry ,AgNOR and PCNA) with quantitative compute processed imaging analysis before intra-arterial infusion chemotherapy and after intra-arterial infusion chemotherapy.
Results : Statistical analysis showed that there were significant differences between the groups (P <0.05). ⑴ There was significant differences in 9 parameters (Prmtr, At ,Dmax, Vv ,Nv , Sv , V, S, Rsv) between before and after intra-arterial infusion chemotherapy of nuclear morphometry . ⑵AgNOR counts of before and after intra-arterial infusion chemotherapy were 6.70± 0.92 and 4.05±0.62. After treatment ,the AgNOR configuration changed greatly . ⑶Positive rates of PCNA before and after intra-arterial infusion chemotherapy were significant difference .
Conclusion : Intera-arterial infusion chemotherapy can inhibit the proliferation of cervical cancer cell . It is recommended that intra-arterial infusion chemotherapy be employed to be one of combined treatments for cervical cancer.
引文
1、 岳天孚，朱雪琼，宫颈癌的术前介入治疗，实用妇产科杂志，2001，17（2）：76—77
2、 Kigawa J,Minagawa Y,et al.The role of neoadjuvant intraarterial infusion chemotherapy with cisplatin and bleomycin for locally advanced cervical cancer.Am J clin oncol(cct)1996,19:225
3、 Zanetta G,Lissoni A,et al.Intense neoadjuvant chemotherapy with cisplatin and epirubicin for advanced or bulky cervical and vaginal adenocarcinoma.Gynecol oncol 1997,64:431
4、 陈春林，梁立治等。介入治疗在中晚期妇科恶性肿瘤中应用的临床研究，中国实用妇科与产科杂志，2000，16（11）：667
5、 申 洪，沈忠英主编，实用生物体视学技术 第1版，广州；中山大学出版社。1991：200-220
6、 Weibel ER.Stereological methods. Vol.l practical methods for biological morphometry.London:Academic press. 1979:68
7、 llan Y,et al. Application of morphometry in tumor pathology.Analyt Quant cytol 1987;9:79
8、 Hall TL, et al. Applications of quantitative microscopy in tumor pathology. Lab Invest 1985;53:5
9、 王伯 ，李玉松等主编，病理学技术，人民卫生出版社，2000：221
10、 许良中，关于核仁组织区（AgNOR）研究工作的标准化方案， 实用肿瘤杂志，1996，11（4）
11、吕卫国,石一复,宫颈癌新辅助化疗前后PCNA表达的研究,实用妇产科杂志,1999,15(3):147-148
12、 左连富，郭建文等，3050例恶性肿瘤DNA倍体分析[J].中华病理学杂志,1992,21:167-168
13、 徐兴钊，李劲松等， 宫颈鳞状细胞癌癌前及癌变病变图像分析， 中华物理医学杂志，1995，17（3）：142-144
14、 焦玉冰，周羡梅等， 子宫颈上皮癌变病变及子宫颈癌的细胞核DNA含量图像分析研究，中华妇产科杂志，2001，36（5）：267-270
15、 Goms,E C.M,Kurver,P.H.J.et al, Morphometric grading Of bladder tumors in comparison with histologic grading by pathologists, Hum Pathol. 1983,14:144-150
16、 Helliwell,T.R, Atkinson,M.W, et al, Morphometric analysis,ploidy and respnonse to chemotherapy in squamous carcinomas of the head and neck. Pathol.Res.Pract. 1989,185:755-759
17、 Partin,A.W, Walsh,A.C, et al, Nuclear morphometry as a predictor of response to therapy in Wilms tumor. A preliminary report, J. Urol,1990,144:952-954
18、 Sherif F.Yacoub,James Shaeffer,et al. Nuclear Morphometry as a Predictor of Response to Neoadjuvant Chemotherapy Plus Radiotherapy in Locally Advanced Cervical Cancer. Gynecologic Oncology,1994,54:327-332
19、 刘履光. 晚期宫颈癌局部动脉灌注化疗的超微结构研究. 武汉医学院学报, 1985,1:38
20、 俞孝庭编著， 肿瘤病理学基础. 第1版. 上海: 上海科学技术出版社. 1986:21-70
21、 Deligdisch L, Carlos Miranda, Joreph Barba, et al. Ovarian Dysplasia: Nuclear Texture analysis. Cancer 1993;72:3253-3257
22、 Schipper NW, Smeulder AWM, Baak JPA. Evaluation of automated estimation of epithelial volume and its prognostic value in ovarian Tumors. Cancer 1989;61:228-233
23、 Derenzim M. Quantity of nuclear silver stained proteins is related to prolterating activity in cancer. Cab Znvest,1990,1:137
Williams MA, Argyrophilic nuclear and nucleolar protein of xenopus laerisocytes identified by lelectrophoresis. Exp Cell
24、 Research, 1982,137:341
25、 Hall PA, Crocker J,et al. Acomparison of nucleolar organizer region staing ﹢ki67 immunostaining in nonflodykics lymphona. Histopathology, 1986,12:375
26、 Anonymous. NoRs-A new method for the pathologist. Cancet,1987,1:1413
27、 吕卫国,陈怀增等, 宫颈癌新辅助化疗后肿瘤组织嗜银核仁组成区变化的研究. 中国实用妇科与产科杂志, 1999,15(10): 599-601
28、 Underwood JCE. Nuclear organizer region as diagnostic discriminals for malignancy. J Pathol, 1988,155:95
29、 张金玲,何秀萍等, AgNOR图像分析在子宫颈鳞癌中的定量研究. 哈尔滨医科大学学报, 1999,33(3):198-200
30、 糜若然, 瞿全新编著. 妇科肿瘤基因诊断与治疗. 人民卫生出版社, 2001:163
31、 Robbins BA, Vega DDL, et al. Immunohistochemical detection of proliferating cell nuclear antigen in solid human malignancies. Arch Pathol Lab Med [J], 1987,111(9):841-845
32、 Terada T, Nakanuma Y,et al. Cell proliferative activity in adenomalous hyperplasia of the liver and small hepatocellular carcinoma. Cancer, 1992,70:591
33、 林志武，晋雯，大肠肿瘤PCNA、AgNOR、DNA及核型态定量研究。中国癌症杂志，2001，11（4）：343-346
34、 Konishi I,Nanbu K,Mandai M,et al. Tumor response to neoadjuvant chemotherapy correlates with the expression of P-glycoprotein and PCNA but not GST-∏ in the tumor cells of cervical carcinoma [J]. Gynecol Oncol,1998,70(3):365-371
35、吕卫国,石一复等,宫颈癌新辅助化疗前后PCNA表达的研究。实用妇产科杂志，1999，15（3）：147-148
36、Ritsuto F, Kentaro T, Manabu K, et al. Decreaase in tumor volume and histologic response to introarterial neoad juvant chemotherapy in patients with cervical and endometrial adenocarcinoma [J]. Gynecol Oncol,1997,65(2):258
37、Astrid M, Masafumi T, Elke F, et al. The pronostic signifisance of proliferition cell nuclear antigen, Epidermal growth Factor Recepter and mdr gene expression in colorectal cancer [J]. Cancer,1993,71(69):2454
38、Giuseppe G ,Andrea C,Caia G, et al. Proliferrating cell nuclear antigen in endometrial carcinoma: pretreatment identification at high-risk patients[J]. Gynecol Oncol, 1996,61(1):16
2、 Kigawa J,Minagawa Y,et al.The role of neoadjuvant intraarterial infusion chemotherapy with cisplatin and bleomycin for locally advanced cervical cancer.Am J clin oncol(cct)1996,19:225
3、 Zanetta G,Lissoni A,et al.Intense neoadjuvant chemotherapy with cisplatin and epirubicin for advanced or bulky cervical and vaginal adenocarcinoma.Gynecol oncol 1997,64:431
4、 陈春林，梁立治等。介入治疗在中晚期妇科恶性肿瘤中应用的临床研究，中国实用妇科与产科杂志，2000，16（11）：667
5、 申 洪，沈忠英主编，实用生物体视学技术 第1版，广州；中山大学出版社。1991：200-220
6、 Weibel ER.Stereological methods. Vol.l practical methods for biological morphometry.London:Academic press. 1979:68
7、 llan Y,et al. Application of morphometry in tumor pathology.Analyt Quant cytol 1987;9:79
8、 Hall TL, et al. Applications of quantitative microscopy in tumor pathology. Lab Invest 1985;53:5
9、 王伯 ，李玉松等主编，病理学技术，人民卫生出版社，2000：221
10、 许良中，关于核仁组织区（AgNOR）研究工作的标准化方案， 实用肿瘤杂志，1996，11（4）
11、吕卫国,石一复,宫颈癌新辅助化疗前后PCNA表达的研究,实用妇产科杂志,1999,15(3):147-148
12、 左连富，郭建文等，3050例恶性肿瘤DNA倍体分析[J].中华病理学杂志,1992,21:167-168
13、 徐兴钊，李劲松等， 宫颈鳞状细胞癌癌前及癌变病变图像分析， 中华物理医学杂志，1995，17（3）：142-144
14、 焦玉冰，周羡梅等， 子宫颈上皮癌变病变及子宫颈癌的细胞核DNA含量图像分析研究，中华妇产科杂志，2001，36（5）：267-270
15、 Goms,E C.M,Kurver,P.H.J.et al, Morphometric grading Of bladder tumors in comparison with histologic grading by pathologists, Hum Pathol. 1983,14:144-150
16、 Helliwell,T.R, Atkinson,M.W, et al, Morphometric analysis,ploidy and respnonse to chemotherapy in squamous carcinomas of the head and neck. Pathol.Res.Pract. 1989,185:755-759
17、 Partin,A.W, Walsh,A.C, et al, Nuclear morphometry as a predictor of response to therapy in Wilms tumor. A preliminary report, J. Urol,1990,144:952-954
18、 Sherif F.Yacoub,James Shaeffer,et al. Nuclear Morphometry as a Predictor of Response to Neoadjuvant Chemotherapy Plus Radiotherapy in Locally Advanced Cervical Cancer. Gynecologic Oncology,1994,54:327-332
19、 刘履光. 晚期宫颈癌局部动脉灌注化疗的超微结构研究. 武汉医学院学报, 1985,1:38
20、 俞孝庭编著， 肿瘤病理学基础. 第1版. 上海: 上海科学技术出版社. 1986:21-70
21、 Deligdisch L, Carlos Miranda, Joreph Barba, et al. Ovarian Dysplasia: Nuclear Texture analysis. Cancer 1993;72:3253-3257
22、 Schipper NW, Smeulder AWM, Baak JPA. Evaluation of automated estimation of epithelial volume and its prognostic value in ovarian Tumors. Cancer 1989;61:228-233
23、 Derenzim M. Quantity of nuclear silver stained proteins is related to prolterating activity in cancer. Cab Znvest,1990,1:137
Williams MA, Argyrophilic nuclear and nucleolar protein of xenopus laerisocytes identified by lelectrophoresis. Exp Cell
24、 Research, 1982,137:341
25、 Hall PA, Crocker J,et al. Acomparison of nucleolar organizer region staing ﹢ki67 immunostaining in nonflodykics lymphona. Histopathology, 1986,12:375
26、 Anonymous. NoRs-A new method for the pathologist. Cancet,1987,1:1413
27、 吕卫国,陈怀增等, 宫颈癌新辅助化疗后肿瘤组织嗜银核仁组成区变化的研究. 中国实用妇科与产科杂志, 1999,15(10): 599-601
28、 Underwood JCE. Nuclear organizer region as diagnostic discriminals for malignancy. J Pathol, 1988,155:95
29、 张金玲,何秀萍等, AgNOR图像分析在子宫颈鳞癌中的定量研究. 哈尔滨医科大学学报, 1999,33(3):198-200
30、 糜若然, 瞿全新编著. 妇科肿瘤基因诊断与治疗. 人民卫生出版社, 2001:163
31、 Robbins BA, Vega DDL, et al. Immunohistochemical detection of proliferating cell nuclear antigen in solid human malignancies. Arch Pathol Lab Med [J], 1987,111(9):841-845
32、 Terada T, Nakanuma Y,et al. Cell proliferative activity in adenomalous hyperplasia of the liver and small hepatocellular carcinoma. Cancer, 1992,70:591
33、 林志武，晋雯，大肠肿瘤PCNA、AgNOR、DNA及核型态定量研究。中国癌症杂志，2001，11（4）：343-346
34、 Konishi I,Nanbu K,Mandai M,et al. Tumor response to neoadjuvant chemotherapy correlates with the expression of P-glycoprotein and PCNA but not GST-∏ in the tumor cells of cervical carcinoma [J]. Gynecol Oncol,1998,70(3):365-371
35、吕卫国,石一复等,宫颈癌新辅助化疗前后PCNA表达的研究。实用妇产科杂志，1999，15（3）：147-148
36、Ritsuto F, Kentaro T, Manabu K, et al. Decreaase in tumor volume and histologic response to introarterial neoad juvant chemotherapy in patients with cervical and endometrial adenocarcinoma [J]. Gynecol Oncol,1997,65(2):258
37、Astrid M, Masafumi T, Elke F, et al. The pronostic signifisance of proliferition cell nuclear antigen, Epidermal growth Factor Recepter and mdr gene expression in colorectal cancer [J]. Cancer,1993,71(69):2454
38、Giuseppe G ,Andrea C,Caia G, et al. Proliferrating cell nuclear antigen in endometrial carcinoma: pretreatment identification at high-risk patients[J]. Gynecol Oncol, 1996,61(1):16