CD44V6在胃癌中的相关研究及其对放射免疫导向手术的评估
摘要
目的 通过研究不同类型胃粘膜上皮、胃癌组织、淋巴结、转移淋巴结、Douglas陷窝腹膜以及种植转移灶中CD44V6的表达情况探讨其作为一项指标在胃癌的分子诊断、分子定界、分子分期及分子预后中应用的可能性并且将之与HE染色结合评价RIGS手术的可行性。方法2001.3.1-2002.3.1，共30例胃镜下及病理证实为胃癌的病人进入治疗组，其中男21例，女9例，年龄范围44-72岁，平均年龄59±8岁，术前采用了~(131)I标记的胃癌单克隆抗体3H_(11)在胃镜下注射入瘤体四周，注射后4-8天进行手术。术中以GDP分别探测胃癌原发灶、切缘、吻合口、肝、脾、各组淋巴结、盆腔等组织，并将探测结果仔细记录。30人均进行了根治性手术。术后将标本组织以及30例正常的胃粘膜组织、50例异型增生组织、良性疾病取材的淋巴结及腹膜各20例的CD44V6的表达情况应用免疫组化方法进行分析及对比。并将其表达情况与血清CEA的检测结果进行比较。结果 (1)正常胃粘膜、良性疾病淋巴结和腹膜不表达CD44V6，而异型增生组织，胃癌组织、转移淋巴结和Douglas腹膜种植转移灶表达CD44V6。而且异型增生与胃癌组织间CD44V6表达强度有显著性差异；(2)CD44V6的表达与胃癌的生长方式、TNM分期、淋巴结转移、脉管浸润和胃癌癌灶浸润深度密切相关；(3)胃癌组织CD44V6的表达率为76.7％，转移淋巴结为88.64％；(4)CD44V6免疫组化与HE结合应用评价RID对胃癌原发灶、切缘、吻合口、转移淋巴结及Douglas陷窝种植转移的准确率分别达到100％，100％，100％，92.43％和83.3％：(5)RID探测结果结合CD44V6免疫组化与HE染色对胃癌TNM分期结果与传统分期方法是有统计学差异的；(6)CD44V6阳性组术后血清CEA阳性率较阴性组高并具有显著性差异。结论 (1)CD44V6与胃癌的发生发展密切相关。CD44V6的表达可以反映胃癌细胞具有较恶的生物学行为。CD44V6可以作为一项指标应用于胃癌的分子诊断、分子定界、分子分期和分子预后中；(2)RIGS可以作为一项可靠的技术应用
安徽医科大学硕士学位论文
于胃癌的治疗中，但仍然存在一定的假阳性和假阴性率，需要进一步的改进。
OhieCtive To discuss the POssibility to aPP1y the CD44V6 prowin as an
index in molecule diagnosis, demarkng, stage and prognosis of gaStric cancer by
comParing with the exPression of CD44V6 in different tyPes of gastric mucosa, lymph
node and PeritOneum of Douglas lacuna. To estimate the feaSibility of RIGS to be
aPPlied in gastric cancer by CD44V6 immunohistochemisty and HE. Methods From
MarCh. 200l to March. 2002, 30 Patients of gastric cancer verifided by gaStrscopy and
hiStology were accepeted in the theraPeutic grouP, 2l were male and 9 were female. The
age ranges frOm 44 to 72 years, the average was 59i8 years. The monocolone of
..,
gastnc carcinoma, 3H11 markd by '3'I was injected thrOugh g8stroscOPy around
cancerous focus PreOpehoon. OPerations were PerfOnned 4 to 8 days later The gaInIna
dectecting Probe was used in the oPeration to deteC the Primp cancer and its
dissemination. The resultS wer reCOrdd wicularly in the PrOCess of laparotmy All
Patients Wrwent radical resection. Serum CEA is also be coInPared betWeen CD44V6
POsitlye and CD4V6 negative grOUs Pre and POst OPeraon. Resube (l) Normal
gaStric mucosa, lyInPh node and perioneum of benign disease dO not exPress CD44V6.
The POsitive rate of CD44V6 exPression in cancer was sitwficantiy higher than tha of
dysPlasia. (2) The exPession of ooV6 was sighficanti cOndated with the gaStric
cancer infiltraive grOwth Pattrw lymPh node metaStasis, vessel invasion, dePth of
invasion and the stage of M. (3) The exPression rate of CD44V6 was 76.7% and
88.64% in Primary focus and lyInPh nodes resPectivey (4) The detechng accurmp of
to to Primeq fOcus, inciSa margins, anastomOtic stomp metastatic lymPh nodes and
inplantaion fOCus in Douglas lacuna was l00%, l00%, l00%, 92.43% and 83.3%
resPeCtively (5) Result of ryM Stag was sighfiCanly different betwen conventional
method and standard method. (6) The POsitive rate of CEA in CD44V6 POsitive grOuP
was significantly higher than that of CD44V6 negative group postoperation, which may indicate that patients of CD44V6 positive expression have a worse prognosis. Conclusion (l)The expression of CD44V6 was correlated with the gastric carcinogenesis and progression. And it can reflect the malignant behavior of tumor cells of gastric cancer. CD44V6 can be used as an indicator to be applied in the molecule diagnosis, demarking and prognosis of gastric cancer. (2)RIGS can be applied in the areas of therapy of gastric cancer as a dependable technology and it should be further improved because of its false positive and negative rate.
安徽医科大学硕士学位论文
于胃癌的治疗中，但仍然存在一定的假阳性和假阴性率，需要进一步的改进。
OhieCtive To discuss the POssibility to aPP1y the CD44V6 prowin as an
index in molecule diagnosis, demarkng, stage and prognosis of gaStric cancer by
comParing with the exPression of CD44V6 in different tyPes of gastric mucosa, lymph
node and PeritOneum of Douglas lacuna. To estimate the feaSibility of RIGS to be
aPPlied in gastric cancer by CD44V6 immunohistochemisty and HE. Methods From
MarCh. 200l to March. 2002, 30 Patients of gastric cancer verifided by gaStrscopy and
hiStology were accepeted in the theraPeutic grouP, 2l were male and 9 were female. The
age ranges frOm 44 to 72 years, the average was 59i8 years. The monocolone of
..,
gastnc carcinoma, 3H11 markd by '3'I was injected thrOugh g8stroscOPy around
cancerous focus PreOpehoon. OPerations were PerfOnned 4 to 8 days later The gaInIna
dectecting Probe was used in the oPeration to deteC the Primp cancer and its
dissemination. The resultS wer reCOrdd wicularly in the PrOCess of laparotmy All
Patients Wrwent radical resection. Serum CEA is also be coInPared betWeen CD44V6
POsitlye and CD4V6 negative grOUs Pre and POst OPeraon. Resube (l) Normal
gaStric mucosa, lyInPh node and perioneum of benign disease dO not exPress CD44V6.
The POsitive rate of CD44V6 exPression in cancer was sitwficantiy higher than tha of
dysPlasia. (2) The exPession of ooV6 was sighficanti cOndated with the gaStric
cancer infiltraive grOwth Pattrw lymPh node metaStasis, vessel invasion, dePth of
invasion and the stage of M. (3) The exPression rate of CD44V6 was 76.7% and
88.64% in Primary focus and lyInPh nodes resPectivey (4) The detechng accurmp of
to to Primeq fOcus, inciSa margins, anastomOtic stomp metastatic lymPh nodes and
inplantaion fOCus in Douglas lacuna was l00%, l00%, l00%, 92.43% and 83.3%
resPeCtively (5) Result of ryM Stag was sighfiCanly different betwen conventional
method and standard method. (6) The POsitive rate of CEA in CD44V6 POsitive grOuP
was significantly higher than that of CD44V6 negative group postoperation, which may indicate that patients of CD44V6 positive expression have a worse prognosis. Conclusion (l)The expression of CD44V6 was correlated with the gastric carcinogenesis and progression. And it can reflect the malignant behavior of tumor cells of gastric cancer. CD44V6 can be used as an indicator to be applied in the molecule diagnosis, demarking and prognosis of gastric cancer. (2)RIGS can be applied in the areas of therapy of gastric cancer as a dependable technology and it should be further improved because of its false positive and negative rate.
引文
1．董志伟，谷铣之．临床肿瘤学．北京：人民卫生出版社，2000：416
2．吴阶平，裘法祖主编．黄家驷外科学(第6版)．北京，人民卫生出版社，2001：1035
3. Xin Y, Grace A, Gallagher MM et al. CD44V6 in gastric carcinoma: a marker of tumor progression. Appl Immunohistochem Mol Morphol 2001;9(2): 138-142
4. Chen GY, Wang DR. The expression and clinical significance of CD44v in human gastric cancers. World J Gastroenterol 2000; 6(1): 125-127
5. Koyama S, Maruyama T, Adachi S. Expression of epidermal growth factor receptor and CD44 splicing variants sharing exons 6 and 9 on gastric and esophageal carcinomas: a two-color flow-cytometric analysis. J Cancer Res Clin Oncol. 1999; 125(1):47-54
6. Haynes BF, Telen MJ, Hale LP et al. CD44—a molecule involved in leukocyte adherence and T-cell activation. Today Immuno Today, 1989; 10: 423-425
7. Haynes BF, Liao HX, Patton KL. The transmembrane hyaluronate receptor (CD44): multiple functions, multipleforms. Cancer Cell, 1991; 3: 347-351
8. Koopman G, Heider KH, Horst E et al. Activated human lymphocytes and aggressive non-Hodgkin's lymphomas express a homologue of the rat metastasis-associated variant of CD44. J Exp Med, 1993, 1 ;177(4): 897-904
9. Thomas L et al. CD44, the hyaluronic acid cell receptor. Its role in neoplastic invasion and metastatic dissemination. Cancer, 1993; 100: 115-118
10. Screaton GR, Bell MV, Jackson DG et al. Genomic structure of DNA encoding the lymphocyte homing receptor CD44 reveals at least 12 alternatively spliced exons. Proc Natl Acad Sci USA, 1992 15; 89(24): 12160-12164
11．吴在德主编．外科学(第五版)，北京：人民卫生出版社，2001：498
12. Tolg C, Hofmann M, Herrlich P et al. Splicing choice from ten variant exons establishes CD44 variability. Nucleic Acids Res, 1993 11; 21(5): 1225-1229
13. Tahara E. Molecular mechanism of human stomach carcinogenesis implicated in Helicobacterpylori infection. Exp Toxicol Pathol 1998 Sep; 50(4-6): 375-8
14. Sairenji T. Epstein-Barr virus (EBV) infection and gastric carcinoma: the approach through EBV infected epithelial cell lines. J Infect Dis 1999 Jun; 52(3):110-2
15. Maeda K, Chung Y, Onoda N, et al。Proliferating cell nuclear antigen labeling index of preoperative biopsy specimens in gastric carcinoma with special reference to prognosis. Cancer, 1994, 73: 528
16．周烨；周决；施达仁等．流式细胞仪检测胃癌中CD44s及CD44v6表达。上海医科大学学报2000；27(1)：48-50
17．张成武，裘华森，邹寿椿等．胃癌PCNA和CD44V6表达的相互关系及其临床意义的研究．中国普通外科杂志。2000；9(4)：318-320
18．赵爱莲；李吉友．胃癌中CD44V6与p53蛋白的表达及其意义．肿瘤防治研究2000：27(1)：18-20
19．范开席，仲伟霞．胃癌中P16、CD44V6的表达及其相关性的研究．实用肿瘤杂志．2000；15(6)：391-395
20. Meyer T, Hart IR。Mechanism of tumour metastasis. Eur J Cancer. 1998; 34: 214
21．吴泰锽主编．肿瘤学新理论与新技术．上海：上海科技教育出版社，1997：134
22．唐镇生主编．分子外科与基因治疗．上海：上海医科大学出版社，1999：76-86
23．刘恭植．现代医学免疫学．南京：江苏科学技术出版社，2001：211
24 张道衡主编．医学分子生物学．北京：北京医科大学出版社，1999：348
25．邓敬兰，王辉生．~(131)I标记单克隆抗体在荷人胃癌裸鼠及胃癌患者体内的定位及免疫显像研究．中华消化杂志：1990，10：262-264
26．[法]阿兰．菲利普著．胡志林译．肿瘤标记．北京：中国医药科技出版社出版，1996：68
27．龚建平．外科细胞分子生物学的现状和趋势．中国实用外科杂志，1999；19：87-89
28．杨璞．肿瘤免疫学．武汉：湖北科学技术出版社，2001：61
29. Thurston MO, Mojzisik CM. History and development of radioimmunoguided surgery. Semin Colon Rectal Surg. 1995; 6:185-191.
30. Martin DT, Hinkle GH, Tuttle S, et al. Intraoperative radioimmunodetection of colorectal tumor with a hand-held radiation detector. Am J Surg. 1985; 150: 672-675
31. O' Dwyer PJ, Mojzisik CM, Hinkle GH, et al. Intraoperative probe-directed immunodetection using a monoclonal antibody. Arch Surg. 1986; 121: 1391-1394
32. Nieroda Ch, Mojziskc, Sordi A et al. Radioimmunoguided surgery in primary colon cancer. Cancer Detect Prey. 1990; 14: 651-656
33. Sickle-Santanello BJ, O' Dwyer PJ, Mojzisik C, et al. Radioimmunoguided surgery using the monoclonal antibody b72.3 in colorectal tumors. Dis Colon Rectum. 1987; 30: 761-764
34．刘宝国，徐光炜，张梅颖，等．胃癌发射免疫导向手术的研究。中华肿瘤杂志，1994，7(4)：284-287
35．潘乐康；吴功侃；张世绵等．放射免疫导向胃癌根治术的临床研究。大连医科大学学报 1997；19(1)：14-17
2．吴阶平，裘法祖主编．黄家驷外科学(第6版)．北京，人民卫生出版社，2001：1035
3. Xin Y, Grace A, Gallagher MM et al. CD44V6 in gastric carcinoma: a marker of tumor progression. Appl Immunohistochem Mol Morphol 2001;9(2): 138-142
4. Chen GY, Wang DR. The expression and clinical significance of CD44v in human gastric cancers. World J Gastroenterol 2000; 6(1): 125-127
5. Koyama S, Maruyama T, Adachi S. Expression of epidermal growth factor receptor and CD44 splicing variants sharing exons 6 and 9 on gastric and esophageal carcinomas: a two-color flow-cytometric analysis. J Cancer Res Clin Oncol. 1999; 125(1):47-54
6. Haynes BF, Telen MJ, Hale LP et al. CD44—a molecule involved in leukocyte adherence and T-cell activation. Today Immuno Today, 1989; 10: 423-425
7. Haynes BF, Liao HX, Patton KL. The transmembrane hyaluronate receptor (CD44): multiple functions, multipleforms. Cancer Cell, 1991; 3: 347-351
8. Koopman G, Heider KH, Horst E et al. Activated human lymphocytes and aggressive non-Hodgkin's lymphomas express a homologue of the rat metastasis-associated variant of CD44. J Exp Med, 1993, 1 ;177(4): 897-904
9. Thomas L et al. CD44, the hyaluronic acid cell receptor. Its role in neoplastic invasion and metastatic dissemination. Cancer, 1993; 100: 115-118
10. Screaton GR, Bell MV, Jackson DG et al. Genomic structure of DNA encoding the lymphocyte homing receptor CD44 reveals at least 12 alternatively spliced exons. Proc Natl Acad Sci USA, 1992 15; 89(24): 12160-12164
11．吴在德主编．外科学(第五版)，北京：人民卫生出版社，2001：498
12. Tolg C, Hofmann M, Herrlich P et al. Splicing choice from ten variant exons establishes CD44 variability. Nucleic Acids Res, 1993 11; 21(5): 1225-1229
13. Tahara E. Molecular mechanism of human stomach carcinogenesis implicated in Helicobacterpylori infection. Exp Toxicol Pathol 1998 Sep; 50(4-6): 375-8
14. Sairenji T. Epstein-Barr virus (EBV) infection and gastric carcinoma: the approach through EBV infected epithelial cell lines. J Infect Dis 1999 Jun; 52(3):110-2
15. Maeda K, Chung Y, Onoda N, et al。Proliferating cell nuclear antigen labeling index of preoperative biopsy specimens in gastric carcinoma with special reference to prognosis. Cancer, 1994, 73: 528
16．周烨；周决；施达仁等．流式细胞仪检测胃癌中CD44s及CD44v6表达。上海医科大学学报2000；27(1)：48-50
17．张成武，裘华森，邹寿椿等．胃癌PCNA和CD44V6表达的相互关系及其临床意义的研究．中国普通外科杂志。2000；9(4)：318-320
18．赵爱莲；李吉友．胃癌中CD44V6与p53蛋白的表达及其意义．肿瘤防治研究2000：27(1)：18-20
19．范开席，仲伟霞．胃癌中P16、CD44V6的表达及其相关性的研究．实用肿瘤杂志．2000；15(6)：391-395
20. Meyer T, Hart IR。Mechanism of tumour metastasis. Eur J Cancer. 1998; 34: 214
21．吴泰锽主编．肿瘤学新理论与新技术．上海：上海科技教育出版社，1997：134
22．唐镇生主编．分子外科与基因治疗．上海：上海医科大学出版社，1999：76-86
23．刘恭植．现代医学免疫学．南京：江苏科学技术出版社，2001：211
24 张道衡主编．医学分子生物学．北京：北京医科大学出版社，1999：348
25．邓敬兰，王辉生．~(131)I标记单克隆抗体在荷人胃癌裸鼠及胃癌患者体内的定位及免疫显像研究．中华消化杂志：1990，10：262-264
26．[法]阿兰．菲利普著．胡志林译．肿瘤标记．北京：中国医药科技出版社出版，1996：68
27．龚建平．外科细胞分子生物学的现状和趋势．中国实用外科杂志，1999；19：87-89
28．杨璞．肿瘤免疫学．武汉：湖北科学技术出版社，2001：61
29. Thurston MO, Mojzisik CM. History and development of radioimmunoguided surgery. Semin Colon Rectal Surg. 1995; 6:185-191.
30. Martin DT, Hinkle GH, Tuttle S, et al. Intraoperative radioimmunodetection of colorectal tumor with a hand-held radiation detector. Am J Surg. 1985; 150: 672-675
31. O' Dwyer PJ, Mojzisik CM, Hinkle GH, et al. Intraoperative probe-directed immunodetection using a monoclonal antibody. Arch Surg. 1986; 121: 1391-1394
32. Nieroda Ch, Mojziskc, Sordi A et al. Radioimmunoguided surgery in primary colon cancer. Cancer Detect Prey. 1990; 14: 651-656
33. Sickle-Santanello BJ, O' Dwyer PJ, Mojzisik C, et al. Radioimmunoguided surgery using the monoclonal antibody b72.3 in colorectal tumors. Dis Colon Rectum. 1987; 30: 761-764
34．刘宝国，徐光炜，张梅颖，等．胃癌发射免疫导向手术的研究。中华肿瘤杂志，1994，7(4)：284-287
35．潘乐康；吴功侃；张世绵等．放射免疫导向胃癌根治术的临床研究。大连医科大学学报 1997；19(1)：14-17