流式细胞术检测肿瘤相关蛋白在人卵巢上皮性肿瘤裸鼠冻融卵巢组织移植中的意义
류식세포술검측종류상관단백재인란소상피성종류라서동융란소조직이식중적의의
Significance of tumor-associated protein detection by flow cytometry in ovarian cryopreservation and transplantation in nude mice bearing human epithelial ovarian cancer
  • 万方数据
    肿瘤研究与临床 종류연구여림상
    CANCER RESEARCH AND CLINIC
    2013年 11期 721-725,729 ,共6页

    朱根海%洪澜%饶若%王圣坦%蔡俊宏%陈春英%姚茂忠%杨舒盈%贺国丽

    주근해%홍란%요약%왕골탄%채준굉%진춘영%요무충%양서영%하국려

    卵巢肿瘤%低温保存%移植%基因,肿瘤%癌旁组织 卵巢腫瘤%低溫保存%移植%基因,腫瘤%癌徬組織
    란소종류%저온보존%이식%기인,종류%암방조직
    Ovarian neoplasms%Cryopreservation%Transplantation%Genes,neoplasm%Paraneoplasm tissue
    目的 通过对人卵巢上皮性肿瘤裸鼠原位移植瘤的癌旁组织相关蛋白的检测,探讨癌旁正常卵巢组织筛选的标准及冻融卵巢组织移植的可行性.方法 将人卵巢上皮性肿瘤OVCAR3细胞于裸小鼠颈背部近腋窝处皮下种植获取瘤源,行卵巢原位移植后,取癌组织、癌旁近端组织、癌旁中段组织、癌旁远端组织及正常裸鼠卵巢组织,采用流式细胞术检测各组织中的细胞角蛋白-7(CK-7)、CA125、p53、Survivin、基质金属蛋白酶-2(MMP-2)、金属蛋白酶组织抑制物-2(T1MP-2)的表达量;分别取全部指标阴性组、CK-7(-)CA125(-)Survivin(-)组、CK-7(+)CA125(+)Survivin(+)组的组织、癌组织和冻融正常裸鼠卵巢组织进行移植,分析各组移植后的癌变率及CA125水平变化,同时检测原位移植后不同病变程度的裸鼠血清CA125水平.结果 从52份人卵巢上皮性肿瘤裸鼠原位移植瘤模型中获取46份(88.5％)活组织检查正常的癌旁残余卵巢组织;仅卵巢种植裸鼠血清CA125水平高于正常裸鼠(P＜0.01),低于卵巢外种植裸鼠(P＜0.05).原位移植瘤组织中CK-7、CA125、p53、Survivin、MMP-2及TIMP-2的表达率分别为93.3 ％(28/30)、93.3 ％(28/30)、86.7％(26/30)、86.7％(26/30)、83.3％(25/30)、83.3％(25/30),癌旁近端组织的表达率分别为70.0％(21/30)、70.0％(21/30)、63.3％(19/30)、63.3％(19/30)、60.0％(18/30)、56.7％(17/30),癌旁中段组织的表达率分别为33.3％(10/30)、30.0％(9/30)、30.0％(9/30)、30.0％(9/30)、26.7％(8/30)、23.3％(7/30),癌旁远端组织的表达率分别为26.7％(8/30)、23.3％(7/30)、26.7％(8/30)、26.7 ％(8/30)、30.0％(9/30)、30.0％(9/30);20份原位移植正常卵巢组织上述指标均为阴性.癌旁近端组织中CK-7、CA125、p53、Survivin、MMP-2及TIMP-2的表达率低于癌组织(P＜0.05),高于癌旁中段组织及癌旁远端组织(P＜0.01);癌旁中段组织及癌旁远端组织表达率差异无统计学意义(P＞0.05).癌旁组织CK-7、CA125、Survivin的强阳性表达率明显高于p53、MMP-2、TIMP-2(P＜ 0.05).全部指标表达阴性组或CK-7(-)CA125(-)Survivin(-)组的组织移植后,未发现癌变,其癌变率及CA125水平低于CK-7(+)CA125(+)Survivin(+)组(P＜0.01,P＜0.05).结论 CK-7、CA125、p53、Survivin、MMP-2及TIMP-2等分子指标的表达向无癌方向呈递减趋势,这些肿瘤相关基因表达全部阴性或主要指标CK-7、CA125、Survivin阴性均可作为筛选癌旁残余正常卵巢组织的标准,流式细胞术可作为筛查残留癌灶及隐匿转移的有效方法;人卵巢上皮性肿瘤裸鼠冻融卵巢组织移植安全、可行.
    목적 통과대인란소상피성종류라서원위이식류적암방조직상관단백적검측,탐토암방정상란소조직사선적표준급동융란소조직이식적가행성.방법 장인란소상피성종류OVCAR3세포우라소서경배부근액와처피하충식획취류원,행란소원위이식후,취암조직、암방근단조직、암방중단조직、암방원단조직급정상라서란소조직,채용류식세포술검측각조직중적세포각단백-7(CK-7)、CA125、p53、Survivin、기질금속단백매-2(MMP-2)、금속단백매조직억제물-2(T1MP-2)적표체량;분별취전부지표음성조、CK-7(-)CA125(-)Survivin(-)조、CK-7(+)CA125(+)Survivin(+)조적조직、암조직화동융정상라서란소조직진행이식,분석각조이식후적암변솔급CA125수평변화,동시검측원위이식후불동병변정도적라서혈청CA125수평.결과 종52빈인란소상피성종류라서원위이식류모형중획취46빈(88.5％)활조직검사정상적암방잔여란소조직;부란소충식라서혈청CA125수평고우정상라서(P＜0.01),저우란소외충식라서(P＜0.05).원위이식류조직중CK-7、CA125、p53、Survivin、MMP-2급TIMP-2적표체솔분별위93.3 ％(28/30)、93.3 ％(28/30)、86.7％(26/30)、86.7％(26/30)、83.3％(25/30)、83.3％(25/30),암방근단조직적표체솔분별위70.0％(21/30)、70.0％(21/30)、63.3％(19/30)、63.3％(19/30)、60.0％(18/30)、56.7％(17/30),암방중단조직적표체솔분별위33.3％(10/30)、30.0％(9/30)、30.0％(9/30)、30.0％(9/30)、26.7％(8/30)、23.3％(7/30),암방원단조직적표체솔분별위26.7％(8/30)、23.3％(7/30)、26.7％(8/30)、26.7 ％(8/30)、30.0％(9/30)、30.0％(9/30);20빈원위이식정상란소조직상술지표균위음성.암방근단조직중CK-7、CA125、p53、Survivin、MMP-2급TIMP-2적표체솔저우암조직(P＜0.05),고우암방중단조직급암방원단조직(P＜0.01);암방중단조직급암방원단조직표체솔차이무통계학의의(P＞0.05).암방조직CK-7、CA125、Survivin적강양성표체솔명현고우p53、MMP-2、TIMP-2(P＜ 0.05).전부지표표체음성조혹CK-7(-)CA125(-)Survivin(-)조적조직이식후,미발현암변,기암변솔급CA125수평저우CK-7(+)CA125(+)Survivin(+)조(P＜0.01,P＜0.05).결론 CK-7、CA125、p53、Survivin、MMP-2급TIMP-2등분자지표적표체향무암방향정체감추세,저사종류상관기인표체전부음성혹주요지표CK-7、CA125、Survivin음성균가작위사선암방잔여정상란소조직적표준,류식세포술가작위사사잔류암조급은닉전이적유효방법;인란소상피성종류라서동융란소조직이식안전、가행.
    Objective To investigate screening criteria of paraneoplastic normal ovarian tissue and the feasibility of human ovarian cryopreservation and transplantation in nude mice by detecting tumorassociated protein.Methods Human epithelial ovarian cancer cell line OVCAR3 were grown in subcutaneous tissue and the tumor tissues were orthotopic implanted.The expressions of CK-7,CA125,p53,Survivin,MMP-2 and TIMP-2 were detected by flow cytometry in tumor tissues,tissues of proximal,median and distal adjacent to tumor,as well as normal ovarian tissues of nude mice.The tissues of group with all negative iudicators,group with negative expressions of CK-7 CA125 Survivin,group with positive expressions of CK-7 CA125 Survivin,cancer tissues and normal ovarian tissues of nude mice were used for ovarian cryopreservation and transplantation.The carcinogenesis rate and CA125 level were analyzed in each group after transplantation.The CA125 level was also detected under different severities after orthotopic implantation in nude mice.Results 46 samples ovarian tissues with normal biopsy were gained from 52 cases of human epithelial ovarian cancer of orthotopic implantation model in nude mice.The CA125 level of only ovarian inoculation group was higher than those of control group (P ＜ 0.01),and lower than those of outside ovarian inoculation group (P ＜ 0.05).The expression rates of CK-7,CA125,p53,Survivin,MMP-2 and TIMP-2 in tumor tissues were 93.3 ％ (28/30),93.3 ％ (28/30),86.7 ％ (26/30),86.7 ％ (26/30),83.3 ％ (25/30),83.3 ％ (25/30) respectively.They were 70.0 ％ (21/30),70.0 ％ (21/30),63.3 ％ (19/30),63.3 ％ (19/30),60.0 ％ (18/30),56.7 ％ (17/30) in proximal tissues adjacent to tumor respectively,and they were 33.3 ％ (10/30),30.0 ％ (9/30),30.0 ％ (9/30),30.0 ％ (9/30),26.7 ％ (8/30),23.3 ％ (7/30) respectively in median tissues adjacent to tumor.In distal tissues adjacent to tumor were 26.7 ％ (8/30),23.3 ％ (7/30),26.7 ％ (8/30),26.7 ％ (8/30),30.0 ％ (9/30),30.0 ％ (9/30) respectively.The expressions of CK-7,CA125,p53,Survivin,MMP-2 and TIMP-2 were all negative in 20 samples normal ovarian tissues adjacent to tumor.The expressions of CK-7,CA125,p53,Survivin,MMP-2 and TIMP-2 in proximal tissues adjacent to tumor were lower than those in tumor tissues (P ＜ 0.05),and higher than those in median and distal tissues adjacent to tumor (P ＜ 0.01).The expressions were no difference between in median and distal tissues (P ＞ 0.05).The strong positive expressions of CK-7,CA125,Survivin were higher than those of p53,MMP-2 and TIMP-2 (P ＜ 0.05).The expressions of CK-7,CA125,p53,Survivin,MMP-2 and TIMP-2 were all negative in 20 samples normal ovarian tissues of nude mice.If all indicators or the main indicators,like CK-7,CA125 and Surrv ivin were not expressed,the tissues did not develop canceration after transplantation.And the carcinogenesis rate and serum CA125 level were lower than that of the group with positive expressions of CK-7,CA125 and Survivin (P ＜ 0.05).Conclusions The expressions of CK-7,CA125,p53,Survivin,MMP-2 and TIMP-2 show decreasing trend to non-cancer direction.Negative expressions of all these tumor-associated genes or the main indicators,CK-7,CA125 and Survivin can be organized as the screening criteria of security of ovarian tissues for transplantation.Flow cytometry can be used as the effective detection method.Ovarian cryopreservation and transplantation in nude mice bearing human epithelial ovarian cancer is feasible and safe.
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