原发性肝癌患者外周血微转移检测的临床意义
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摘要
目的:建立检出原发性肝癌(PHC)患者外周血中微转移的方法,并探讨其临床意义。
     方法:采集50例PHC患者,40例肝良性疾病(10例肝血管瘤,10例肝囊肿,10例肝硬化,10例慢性肝炎)患者,20例肝转移肿瘤患者(直肠癌肝转移10例,胃癌肝转移10例),30例健康志愿者的外周血。用RT-PCR方法检测外周血中AFPmRNA和CD44v6mRNA的表达。
     结果:50例PHC患者外周血中AFPmRNA和CD44v6mRNA检出率分别为50%(25/50)和58%(29/50)。74%(37/50)的患者至少一种标志物阳性。AFPmRNA在肝炎肝硬化患者外周血中的检出率为15%(3/20),转移性肝癌、肝血管瘤、肝囊肿患者及健康献血员中均未检出。CD44v6mRNA在转移性肝癌患者外周血中的检出率为45%(9/20),肝炎肝硬化、肝血管瘤、肝囊肿患者及健康献血员中均未检出。PHC患者外周血中AFPmRNA检出率与血清AFP水平,肿瘤大小,门静脉癌栓,肝内转移无相关性(p>0.05)。与肝外转移密切相关(p<0.05)。CD44v6mRNA检出率与肿瘤大小,门静脉癌栓,肝内外转移密切相关(p<0.05),而与血清AFP水平无相关性(p>0.05)。
     结论:在PHC患者外周血中联合检测AFPmRNA和CD44v6mRNA的表达,有助于提高微转移检测的灵敏度和特异性,CD44v6mRNA预测肝癌转移、
Objective: To establish the molecular method to detect peripheral blood micrometsastasis in patients with Primary hepatocellular carcinoma(PHC),and discuss its clinical sigificance.
     Methods: A sensitive RT-PCR system was set up to detect AFPmRNA and CD44v6mRNA in the peripheral blood of 50 PHC patients, 10 hepatic hemangioma patients ,10 hepatic cyst patients,10 hepatocirrhosis patients,10 chronic hepatitis patients, 20 patients with metastatic liver cancer, and 30 healthy volunteers.
     Results: Of the PHC patients, expression of AFPmRNA and CD44v6mRNA was positive respectively in 50%(25/60) and 58%(29/50) of the peripheral bloods.while 37(74% )PHC patients were positive for at least 1 marker. With regard to controls, 3 of 20 samples(15%)from group hepatitis and cirrhosis showed AFPmRNA,but none of samples from other control group showed AFPmRNA. And 9 of 20(45%)samples from patients with metastatic liver cancer showed CD44v6mRNA in their peripheral blood,but groups of hepatitis and cirhosis,hepatic hemangioma,hepatic
引文
[1] Hayashi N,Ito I,Yanagisawa A,et al.Genetic diagnosis of lymph-node metastases in colorectal cancer[J]. Lancetm,1995,345:1257-1259.
    [2] Ghossein RA,Osman I, Bhattacharya S, et al. Detection of prostatic specific membrane antigen messenger RNA using immunobead reverse transcriptase polymerase chain reaction[J]. Diagn Mol Pathol,1999,8:59-65.
    [3] 黄东胜,傅佩芬,郑树森,等. 肝癌病人外周血及骨髓 AFP mRNA 的检测及意义[J].中华肝胆外科杂志,2002.03.28; 8(3): 154-158.
    [4] 彭安邦,石巍,胡四海,等. CD44v6 在胃癌中的表达及其与幽门螺杆菌感染的关系[J]。癌症杂志,2003,22(11):1184-1187.
    [5] Mutsuko M,Naoshi N, et al.Postoperative detection of a-fetoprotein mRNA in blood as a predictor for metastatic recurrence of hepatocellular carcinoma[J]. Gastroenterol Hepatol,2001,16(4):445-451.
    [6] Miyamoto,-A; Fujiwara,-Y; Sakon,-M;et al. Development of a multiple-marker RT-PCR assay for detection of micrometastases of hepatocellular carcinoma[J]. Dig-Dis-Sci. 2000 Jul; 45(7): 1376-82
    [7] Kedda MA, Kew MC, Skelton M, et al. Nonspecific messenger RNA of a-fetoprotein in peripheral blood in detecting early spread of hepatocellular carcinoma in black Africans[J]. Gastroenterol Hepatol,1998,13(6):885-891.
    [8] Rui JA,Wang SB,Chen SG,et a1.Serial Therapies Oriented by Surgery for Large Primary Liver Carcinoma [J].Hepatobiliary Pancreat Dis Int,2002,1(3):411-415.
    [9] Liu Y,Wu Mc,Oin Gx,et a1.Nested RT—PCR in De tection of Blod AFPmRNA in Animal Model of Human Hepatoeellular Carcinoma[J]. Hepatobiliary Pancreat Dis Int,2003,2(3):414-417.
    [10] Liu Y,Zhang BH,Qian GX,et al. Expression of AFPmRNA in blood of patients with recurrent hepatocellular carcinoma[J].Hepatobiliary Pancreat Dis Int. 2003 May;2(2):274-7.
    [11] Kienle P; Weitz J; Klaes R, et al. Detection of isolated disseminated tumor cells in bone marrow and blood samples of patients with hepatocellular carcinoma[J]. Arch-Surg. 2000 Feb; 135(2): 213-218。
    [12] Chuang CK,Liao SK, et al. Differential expression of CD44 variant isoforms by cell lines and tissue specimens of transitional cell carcinomas[J].Anticancer Res. 2003 Nov-Dec;23(6C):4635-9.
    [13] Gao C,Guo H,Downey L,et al. Osteopontin-dependent CD44v6 expression and cell adhesion in HepG2 cells.Carcinogenesis[J]. 2003 Dec;24(12):1871-8. Epub 2003 Aug 29.
    [14] Yamada Y,Itano N,Narimatsu H,et al. CD44 variant exon 6 expressions in colon cancer assessed by quantitative analysis using real time reverse transcriptase-polymerase chain reaction[J]. Oncol Rep.2003 Nov-Dec; 10(6):1919-24.
    [15] Indinnimeo M,Cicchini C,Giarnieri E,et al. Evaluation of CD44 variant 6 expression and clinicopathological factors in pulmonary metastases from colon carcinoma[J].Oncol Rep. 2003 Nov-Dec;10(6):1875-7.
    [16] Gulmann C,Grace A,Leader M,et al. CD44v6: a potential marker of malignant transformation in intestinal metaplasia of the stomach? An immunohistochemical study using tissue microarrays[J].Eur J Gastroenterol Hepatol. 2003 Sep;15(9):981-6.
    [17] Yamaguchi A,Goi T,Yu J,et al. Expression of CD44v6 in advanced gastric cancer and its relationship to hematogenous metastasis and long-term prognosis[J].J Surg Oncol. 2002 Apr;79(4):230-5.
    [18] Niu Y, Liu F,Zhou Z,et al. Expression of CD44V6 and PCNA in squamous cell carcinomas[J]. Chin Med J (Engl). 2002 Oct;115(10):1564-8.
    [19] Endo K; Terada T, et al. Protein expression of CD44(standard and variant isoforms)in hepatocellular carcinoma: relationships with tumor grade, clinicopathologic parameters, p53 expression, and patient survival[J]. J-Hepatol. 2000 Jan; 32(1): 78-84。
    [20] 邵宇,黄颖,黄建富. E-钙粘附蛋白,CD44V6在原发性肝细胞癌中表达的研究[J].肝胆外科杂志, 2002.10.25; 10(5): 388-389.
    [21] 谷化平,周翠玲,倪灿荣. 原发性肝细胞癌中CD44v6和nm23H1基因的转录表达及临床意义[J]. 临床肝胆病杂志,2002.06.20; 18(3): 159-160.
    [22] Yan X,Yanping W,Sumin Z,et al. Relationship between CD44v6 expression and metastatic potential and prognosis in gastric carcinoma[J].Chin Med Sci J. 2000 Jun;15(2):128.
    [23] Masson D,Denis MG,Lustenberger P, et al.Limitations of CD44v6 amplification for the detection of tumour cells in the blood of colorectal cancer patients[J].Br J Cancer. 2000 Apr;82(7):1283-9.
    [1] 吴天鸣,杨翔,贺强,等.肝移植治疗原发性肝癌的探讨[J].临床外科杂志,2002,11;10(6):371-372.
    [2] 郑树森,吴健,黄东胜,等.肝脏移植在治疗肝脏恶性肿瘤中的作用[J].中华肝胆外科杂志 2002.05.28;8(5):265-267.
    [3] Schlitt HJ, Neipp M,et al.Recurrence patterns of hepatocellular and fibrolamellar carcinoma after liver transplantation[J]. J-Clin-Oncol. 1999. Jan,17(1): 324-31.
    [4] Castroagudin JF,Gonzalez-Quintela,-A;et al.Bilateral adrenal metastases from hepatocellular carcinoma after liver transplantation[J]. Hepatogastroenterology.2002.Jan-Feb;49(43):249-51.
    [5] Roayaie S,Fricher JS,Emre SH,et al.Long-term results with multimodal adjuvant therapy and liver transplantation for the treatment of hepatocellular carcinomas larger than 5 centimeters[J].Ann Surg.2002,235:533-539.
    [6] Jonas S, Bechstein WO, Steinmuller T, et al.Vascular invasion and histopathologic grading determine outcome after liver transplantation for hepatocellular carcinoma in cirrhosis[J].Hepatology.2001,33:1080–1086.
    [7] Yao FY, Bass NM, Nikolai B, et al.Liver transplantation for hepatocellular carcinoma: analysis of survival according to the intention-to-treat principle and dropout from the waiting list[J]. Liver Transpl.2002,8:873–883.
    [8] Esnaola,-N-F;Lauwers,-G-Y;et al. Predictors of microvascular invasion in patients with hepatocellular carcinoma who are candidates for orthotopic liver transplantation[J].J-Gastrointest-Surg.2002 Mar-Apr;6(2):224-32;discussion 232.
    [9] Henri,Pietro, E,et al.Liver Transplantation for Hepatocellular Carcinoma[J]. Seminars in Liver Disease,1999 , 19(3) :311-322.
    [10] 扬甲梅,温增庆.肝移植术治疗原发性肝癌[J].肝胆外科杂志,1999.7(3):166-167.
    [11] Kienle p,Weitz J,Klaes R,et al.Detection of isolated disseminated tumor cells in bone marrow and vlood samples of patients with hepatocellular carcinoma[J].Arch surg.2000,135(2):213.
    [12] 王槐志,董家鸿,王曙光,等。肝移植治疗原发性肝癌[J],实用临床医药杂志,2003.2;7(1):27-29.
    [13] Hemming AW,Cattral MS,Reed AI,et al.Liver Transplantation for Hepatocellular Carcinoma[J].Analls of surgery.2001, 233(5):652-659.
    [14] Salizzoni M,Romagnoli R,et al.Microscopic vascular invasion detected by anti-CD34 immunohistochemistry as a predictor of recurrence of hepatocellular carcinoma after liver transplantation[J].Transplantation.2003,76(5): 844-8.
    [15] Freise CE,Ferrell L,Liu T, et al.Effect of systemic cyclosporine on tumor recurrence after liver transplantation in a model of hepatocellular carcinoma[J].Transplantation.1999,67(4):510-513.
    [16] Vivarelli M,Bellusci R,Cucchetti A,et al.Low recurrence rate of hepatocellular carcinoma after liver transplantation : better patient selection or lower immunosuppression?[J]. transplantation .2002, 74(12):1746-1751.
    [17] Dumortier J,Lombard-Bohas C,et al.Needle tract recurrence of hepatocellular carcinoma after liver transplantation[J].Gut.2000,47(2): 301.
    [18] Iwatsuki S,Dvorchik I,Marsh JW,et al.Liver transplantation for hepatocellular carcinoma:a proposal of a prognostic scoring system[J].J Am coll surg,2000,191: 389-394.
    [19] Mutsuko M,Naoshi N, et al.Postoperative detection of a-fetoprotein mRNA in blood as a predictor for metastatic recurrence of hepatocellular carcinoma[J].J Gastroenterol Hepato.2001,16:445-451.
    [20] 牛瑞芳,杨毅.肝细胞癌在血中微小转移的检测[J].中华消化杂志,2002.09.20; 22(9): 543-545
    [21] 刘允怡.肝移植在肝癌治疗中的地位与评价 [J] .中华肝胆外科杂志,2003.05.28;9(5):265-267.
    [22] Kaihara S,Kiuchi T,Ueda M,et al. Living-donor liver transplantation for hepatocellular carcinoma[J], transplantation.2003,75(3):S37-S40.
    [23] Stuart J,Challenging Choices : Liver Transplantation for Hepatocellular Carcinoma[J].Analls of surgury.2004, 239(2):160-161.
    [24] Belghiti J,Cortes A,Abdalla EK,et al.Resection Prior to Liver Transplantation for Hepatocellular Carcinoma[J].Analls of surgery.2003, 238(6):885–893.
    [25] Adam R,Azoulay D,et al. Liver resection as a bridge to transplantation for hepatocellular carcinoma on cirrhosis: a reasonable strategy?[J]. Ann-Surg. 2003 Oct;238(4): 508-18;discussion 518-519.
    [26] Adam R,Azoulay D,et al. Liver resection as a bridge to transplantation for hepatocellular carcinoma on cirrhosis: a reasonable strategy?[J]. Ann-Surg. 2003 Oct;238(4): 508-18;discussion 518-519.
    [27] Graziadei I W, Sandmueller H,et al.Chemoembolization followed by liver transplantation for hepatocellular carcinoma impedes tumor progression while on the waiting list and leads to excellent outcome[J]. Liver-Transpl.2003 Jun;9(6):557-63.
    [28]肖开银,彭民浩,黎乐群,等。肝移植在小肝癌治疗中的地位[J],肝胆外科杂志2003.8;11(4):259-261.
    [29] Fontana RJ,Hamidullah H,Nghiem H,et al.Percutaneous radiofrequency thermal ablation of hepatocellular carcinoma:a safe and effective bridge to liver transplantation[J]. Liver Transpl.2002,8:1165-1174.
    [30] Roayaie S,Fricher JS,Emre SH,et al.Long-term results with multimodal adjuvant therapy and liver transplantation for the treatment of hepatocellular carcinomas larger than 5 centimeters[J].Ann Surg.2002,235:533-539.
    [31] 陈规划,陆敏强,蔡常洁,等. 肝癌肝移植术后个体化化疗疗效初步分析[J].中华外科杂志, 2004,42(17).1040-1043.
    [32] 董家鸿,肝癌肝移植[J],中华肝脏病杂志.2002.2;10(1):61-62.
    [33] 杨世忠,董家鸿.肝细胞癌微转移检测的临床意义及研究进展[J].中华外科杂志2003.3;41(3):228-230.
    [1] Ghossein RA,Osman I, Bhattacharya S, et al. Detection of prostatic specific membrane antigen messenger RNA using immunobead reverse transcriptase polymerase chain reaction[J]. Diagn Mol Pathol,1999,8:59-65.
    [2] 朱广迎, 陈杰, 朱斌,等.巢式 RT-PCR 检测肝癌患者外周血微量转移癌细胞[J].徐州医学院学报,2001.01.15; 21(1): 1-3.
    [3] Peter K, Alfred Z, Ralf L,et al. Heterogenous expression of MEGA-A Genes in occult disseminated tumor cells: diagnosis of micrometastatic disease[J]. Cancer Res, 2002,62: 251-261.
    [4] Miyamoto A, Nagano H, Sakon M, et al. Clinical application of quantitative analysis for detection of hematogenous spread of hepatocellular carcinoma by real-time PCR[J].Int J Oncol,2001,18:527-532.
    [5] 吴晓凤,林芷英,樊嘉等.外周血甲胎蛋白 mRNA 定量与裸鼠肝癌术后复发转移的关系[J].中华肝脏病杂志,2002,06,10(3):189-192.
    [6] 黄瑜峰, Oberneder R, Piesenberg R,等.肿瘤细胞在骨髓中播散的自动显微图像研究[J].广西医科大学学报,2000.04.25; 17(2): 169-172.
    [7] 赵秀兰, 宋文静,刘易欣,等.局域淋巴结微转移癌的免疫组织化学检测及意义[J],天津医科大学学报.2003.09.15; 9(3): 334-336.
    [8] Mutsuko M,Naoshi N, et al .Postoperative detection of a-fetoprotein mRNA in blood as a predictor for metastatic recurrence of hepatocellular carcinoma[J] .J Gastroenterol Hepato,2001,16:445-451.
    [9] Kienle P,Weitz J, Klaes R , et al . Detection of isolated disseminated tumor cells in bone marrow and blood samples of patients with hepatocellular carcinoma[J]. Arch-Surg. 2000 Feb; 135(2): 213-218.
    [10] 周京旭, 杨希山, 袁爱力等.应用巢式 RT-PCR 检测原发性肝细胞癌患者外周血中的微转移[J].中国肿瘤临床与康复,2000.01.20; 7(1): 8-10.
    [11] 黄东胜,傅佩芬,郑树森,等.肝癌病人外周血及骨髓 AFP mRNA 的检测及意义[J].中华肝胆外科杂志,2002.03.28; 8(3): 154-158.
    [12] Wu GX, Lin YM, Zhou TH, et al . Significant down-regulation of alpha-albumin in human hepatoma and its implication[J]. Cancer Lett, 2000,160:229-236.
    [13] 牛瑞芳,杨毅,魏熙胤,等.肝细胞癌在血中微小转移的检测[J].中华消化杂志,2002.09.20; 22(9): 543-545.
    [14] 陶则东, 程炳立,王海滨,等.肝癌患者红细胞膜 CD44s 表达的变化及临床意义[J].中国医学检验杂志,2002.10.28; 3(5): 303-305.
    [15] Endo K, Terada T, et al .Protein expression of CD44(standard and variant isoforms)in hepatocellular carcinoma: relationships with tumor grade, clinicopathologic parameters, p53 expression, and patient survival[J]. J-Hepatol. 2000 Jan; 32(1): 78-84.
    [16] 管小琴,徐曼,李圆圆,等.CD44 与肝细胞癌的分化和血管浸润[J].重庆医科大学学报, 2002.09.15; 27(3): 247-249.
    [17] 邵宇,黄颖,黄建富. E-钙粘附蛋白,CD44V6在原发性肝细胞癌中表达的研究[J].肝胆外科杂志, 2002.10.25; 10(5): 388-389.
    [18] 谷化平,周翠玲,倪灿荣. 原发性肝细胞癌中 CD44v6 和 nm23H1 基因的转录表达及临床意义[J]. 临床肝胆病杂志,2002.06.20; 18(3): 159-160.
    [19] Miyamoto A, Fujiwara Y, Sakon M, et al . Development of a multiple-marker RT-PCR assay for detection of micrometastases of hepatocellular carcinoma[J]. Dig-Dis-Sci. 2000 Jul; 45(7): 1376-82.
    [20] Kobayashi Y, Higashi T,Nouso K, et al . Expression of MEGA,GAGE and BAGE genes in human liver diseases:utility as molecular markers for hepatocellular carcinoma[J]. J Hepatol,2000,32:612-617.
    [21] Tatsuma T, Goto S, Kitano S, et al . Telomerase activity in peripheral blood for diagnosis of hepatoma[J]. J-Gastroenterol-Hepatol. 2000 Sep; 15(9): 1064-70.
    [22] Park NH, Chung YH, Youn KH, et al . Close correlation of p53 mutation to microvascular invasion in hepatocellular carcinoma[J]. J-Clin-Gastroenterol. 2001 Nov-Dec; 33(5): 397-401.
    [23] 张延龄.微转移的生物学行为和临床意义[J],国外医学外科学分册,2001,28(5):267-268
    [24] 周京旭, 杨希山, 袁爱力,等.手术和 TAE 术前后外周血肝癌播散细胞的变化及随访观察[J].临床消化病杂志,2000.10.20; 12(5): 207-210.
    [25] 周京旭, 杨希山,袁爱力,等.TAE 术前后外周血肝癌播散细胞的变化及随访观察[J].现代消化及介入诊疗,2000.10.15; 5(3): 20-23.
    [26] 刘扬,吴孟超,钱光相,等.联合治疗复发性肝癌血液循环性肝癌细胞的变化及意义[J].中华肝脏病杂志,2001,9:40-41.
    [27] 金成兵, 伍烽, 王智彪,等. 高强度聚焦超声治疗前后肝癌患者外周血AFPmRNA 的变化[J].中国超声医学杂志,2003.11.16; 19(11): 838-841.

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