前列腺酸性磷酸酶体外诱导结肠癌特异性CTLs的研究
|
摘要
结肠癌是胃肠道常见的恶性肿瘤之一,每年有大量患者死于此病,至今仍缺乏有效的早期诊断方法。近20年我国尤其大城市的结肠癌发病率明显上升。由于结肠癌患者初期很少有症状,确诊时60%-70%的结肠癌已属晚期,故死亡率较高。早期结肠癌患者经治疗后的生存率可达90%。因此寻找早期诊断方法,探索有效的综合疗法是降低结肠癌死亡率的关键。
自1938年起,前列腺酸性磷酸酶(prostatic acid phosphatase, PAP)便作为前列腺癌肿瘤标志物。多个报道提示在前列腺和前列腺癌之外的其他组织也有PAP的表达,因此它并不是绝对的组织特异性标志物。
本实验利用RT-PCR法及Western blot法对PAP在结肠癌细胞中的表达水平进行检测,利用免疫组织化学方法对PAP在结肠癌组织中的PAP表达水平进行检测;利用ELISA法检测结肠癌患者和健康人血浆中抗PAP表位肽的抗体水平,利用ELISpot法检测结肠癌及健康人PBMCs中分泌PAP多肽特异性IFN-γ的细胞数量,利用~(51)Cr释放法对PAP表位肽体外诱导产生的细胞毒性T淋巴细胞(cytotoxic T lymphocytes,CTLs)的细胞毒性进行测定,从而为结肠癌的诊断和治疗提供一些实验依据。
Colon cancer is the most common malignant tumor of the gastrointestinal tract, which has been causing many people dead per year. In the recent 20 years in China, especially in the big cities, incidence has been obviously increased. Colon cancer almost has no early special symptoms. The developed symptoms are often changed bowel evacuation habit and excrement feature, abdominal pain and bowel obstruction symptoms. Due to the rapid progress, concealment, initial few symptoms, the colon cancer is still lack of effective early diagnostic methods. When diagnosed, 60%～70% of the colon cancer have been late process. Therefore the mortality rate is very high. While for the early colon cancer patients after treatment, the survival rate can reach 90%. Therefore the early diagnosis methods and effective combination therapy is the key to reduce colorectal cancer mortality. Improving the level of early diagnosis and performing the early treatment are important to improve the prognosis.
Prostatic Acid Phosphatase (PAP) is an enzyme secreted by prostate cancer epithelial cells. It is the main source for the APC. It is an isozyme coming from the Cytolysosome of the prostate cancer epithelial cells. Human prostate is the main source for the enzyme. As the prostate cancer markers, concentration measurement for serum PAP is widely used in the prostate cancer diagnosis and the monitoring for the treatment. Compared with prostate-specific antigen (PSA), PAP is a more accurate indicator for tumor micrometastases. In addition, the PAP is the immune therapy target for the metastatic non-androgen dependence prostatic carcinoma. PAP was once considered as the prostate specific tumor markers. Several articles report that PAP also exists in non-prostate organ. It seems that PAP is not an prostate-specific tumor marker. In previors study, We have confirmed that the positive expression of PAP in the stomach, ovarian and breast cancer. PAP is the new immune therapy target for the stomach, ovarian and breast epithelial tissue cancer. In this study, the PAP expression in colon cancer cell lines and colon cancer tissue and the possibility of PAP as a immunotherapy target of colon cancer were investigated .
(1)The expression of PAP in colon cancer cell lines
1)The expression of PAP mRNA in Colon carcinoma cell lines
The expression of PAP mRNA in various colon carcinoma cell lines was investigated by the RT-PCR method. Total RNA was isolated from 1×10~7 colon carcinoma cells, cDNA was synthesized by RT-PCR. PAP cDNA was detected by PCR amplification using a set of oligonucleotide primers specific to PAP. The mRNA expression of PAP was detected in 3(colo201.colo205.colo320)of 5 colon adenocarcinoma cell lines(colo201, colo 205, colo 320, HCT116 and LoVo ).
2)The expression of PAP protein in colon carcinoma cell lines
The expression of the PAP protein in colon carcinoma cell lines was examined by Western blot analysis. The cytoplasmic protein of different colon carcinoma cell lines were extracted according to the manual. The protein was separated by SDS-PAGE. The proteins in the acrylamide gel were blotted to a Hybond-polyvinylidene difluoride membraneand probed with Anti-PAP mouse monoclonal antibody. The PAP protein was found to be expressed in colo201,colo205 and colo320.
3)Expression of PAP in colon cancer tissues
The PAP expression of colon adenocarcinoma tissues were examined by immunohistochemical staining. As a result, colon cancer tissues were positive for PAP.
(2)Induction of Peptide-Specific CTLs and Assay of Cytotoxicity
1)HLA-A2-restricted PAP epitope peptide
18 PAP epitope peptides which have the binding motif to HLA-A2 molecules, were required from the amino acid sequence of PAP through software analysis. All peptides were dissolved with dimethyl sulfoxide at a concen-tration of 10 mg/mL.
2)Detection of anti- PAP peptide antibody
Anti- PAP peptide antibody(IgG) in the plasma of colon cancer patients and healthy donors were examined by ELISA analysis. High level anti- PAP_(135-143)、PAP_(112-120)、PAP_(33-41)、PAP_(201-210) or PAP_(196-205) IgG could be found in the plasma of most colon cancer patients.
3)Induction of PAP peptide-Specific IFN-γsecreting CTLs
PBMCs of HLA-A2~+ cancer patients or HLA-A2~+ healthy donors were incubated with PAP_(135-143)、PAP_(112-120)、PAP_(33-41)、PAP_(201-210) or PAP_(196-205) peptide respectively. The IFN-γsecretion were examined by the ELISPOT test. It is showed that peptide-specific IFN-γsecretion could be induced by PAP_(112-120) or PAP_(201-210) peptide.
4)~(51)Cr release assay of Cytotoxicity
The cytotoxicity of PAP_(112-120) or PAP_(201-210) peptide-specific CTLs was examined by ~(51)Cr-release assay. As a result, the PAP_(112-120)-specific CTLs lysed the PAP+/HLA-A2+ colon cancer cell colo 205 specificly.
In conclusion, PAP mRNA and PAP protein were revealed to be expressed in colon cancer cell lines. PAP protein were positive in colon cancer tissue. PAP112-120 peptide-specific CTLs induced from colon cancer patients lysed the PAP+/HLA-A2+ colon cancer cell colo 205 specificly. HLA-A2-restricted PAP112-120 peptide could be used in the immunotherapy of colon cancer.
自1938年起,前列腺酸性磷酸酶(prostatic acid phosphatase, PAP)便作为前列腺癌肿瘤标志物。多个报道提示在前列腺和前列腺癌之外的其他组织也有PAP的表达,因此它并不是绝对的组织特异性标志物。
本实验利用RT-PCR法及Western blot法对PAP在结肠癌细胞中的表达水平进行检测,利用免疫组织化学方法对PAP在结肠癌组织中的PAP表达水平进行检测;利用ELISA法检测结肠癌患者和健康人血浆中抗PAP表位肽的抗体水平,利用ELISpot法检测结肠癌及健康人PBMCs中分泌PAP多肽特异性IFN-γ的细胞数量,利用~(51)Cr释放法对PAP表位肽体外诱导产生的细胞毒性T淋巴细胞(cytotoxic T lymphocytes,CTLs)的细胞毒性进行测定,从而为结肠癌的诊断和治疗提供一些实验依据。
Colon cancer is the most common malignant tumor of the gastrointestinal tract, which has been causing many people dead per year. In the recent 20 years in China, especially in the big cities, incidence has been obviously increased. Colon cancer almost has no early special symptoms. The developed symptoms are often changed bowel evacuation habit and excrement feature, abdominal pain and bowel obstruction symptoms. Due to the rapid progress, concealment, initial few symptoms, the colon cancer is still lack of effective early diagnostic methods. When diagnosed, 60%～70% of the colon cancer have been late process. Therefore the mortality rate is very high. While for the early colon cancer patients after treatment, the survival rate can reach 90%. Therefore the early diagnosis methods and effective combination therapy is the key to reduce colorectal cancer mortality. Improving the level of early diagnosis and performing the early treatment are important to improve the prognosis.
Prostatic Acid Phosphatase (PAP) is an enzyme secreted by prostate cancer epithelial cells. It is the main source for the APC. It is an isozyme coming from the Cytolysosome of the prostate cancer epithelial cells. Human prostate is the main source for the enzyme. As the prostate cancer markers, concentration measurement for serum PAP is widely used in the prostate cancer diagnosis and the monitoring for the treatment. Compared with prostate-specific antigen (PSA), PAP is a more accurate indicator for tumor micrometastases. In addition, the PAP is the immune therapy target for the metastatic non-androgen dependence prostatic carcinoma. PAP was once considered as the prostate specific tumor markers. Several articles report that PAP also exists in non-prostate organ. It seems that PAP is not an prostate-specific tumor marker. In previors study, We have confirmed that the positive expression of PAP in the stomach, ovarian and breast cancer. PAP is the new immune therapy target for the stomach, ovarian and breast epithelial tissue cancer. In this study, the PAP expression in colon cancer cell lines and colon cancer tissue and the possibility of PAP as a immunotherapy target of colon cancer were investigated .
(1)The expression of PAP in colon cancer cell lines
1)The expression of PAP mRNA in Colon carcinoma cell lines
The expression of PAP mRNA in various colon carcinoma cell lines was investigated by the RT-PCR method. Total RNA was isolated from 1×10~7 colon carcinoma cells, cDNA was synthesized by RT-PCR. PAP cDNA was detected by PCR amplification using a set of oligonucleotide primers specific to PAP. The mRNA expression of PAP was detected in 3(colo201.colo205.colo320)of 5 colon adenocarcinoma cell lines(colo201, colo 205, colo 320, HCT116 and LoVo ).
2)The expression of PAP protein in colon carcinoma cell lines
The expression of the PAP protein in colon carcinoma cell lines was examined by Western blot analysis. The cytoplasmic protein of different colon carcinoma cell lines were extracted according to the manual. The protein was separated by SDS-PAGE. The proteins in the acrylamide gel were blotted to a Hybond-polyvinylidene difluoride membraneand probed with Anti-PAP mouse monoclonal antibody. The PAP protein was found to be expressed in colo201,colo205 and colo320.
3)Expression of PAP in colon cancer tissues
The PAP expression of colon adenocarcinoma tissues were examined by immunohistochemical staining. As a result, colon cancer tissues were positive for PAP.
(2)Induction of Peptide-Specific CTLs and Assay of Cytotoxicity
1)HLA-A2-restricted PAP epitope peptide
18 PAP epitope peptides which have the binding motif to HLA-A2 molecules, were required from the amino acid sequence of PAP through software analysis. All peptides were dissolved with dimethyl sulfoxide at a concen-tration of 10 mg/mL.
2)Detection of anti- PAP peptide antibody
Anti- PAP peptide antibody(IgG) in the plasma of colon cancer patients and healthy donors were examined by ELISA analysis. High level anti- PAP_(135-143)、PAP_(112-120)、PAP_(33-41)、PAP_(201-210) or PAP_(196-205) IgG could be found in the plasma of most colon cancer patients.
3)Induction of PAP peptide-Specific IFN-γsecreting CTLs
PBMCs of HLA-A2~+ cancer patients or HLA-A2~+ healthy donors were incubated with PAP_(135-143)、PAP_(112-120)、PAP_(33-41)、PAP_(201-210) or PAP_(196-205) peptide respectively. The IFN-γsecretion were examined by the ELISPOT test. It is showed that peptide-specific IFN-γsecretion could be induced by PAP_(112-120) or PAP_(201-210) peptide.
4)~(51)Cr release assay of Cytotoxicity
The cytotoxicity of PAP_(112-120) or PAP_(201-210) peptide-specific CTLs was examined by ~(51)Cr-release assay. As a result, the PAP_(112-120)-specific CTLs lysed the PAP+/HLA-A2+ colon cancer cell colo 205 specificly.
In conclusion, PAP mRNA and PAP protein were revealed to be expressed in colon cancer cell lines. PAP protein were positive in colon cancer tissue. PAP112-120 peptide-specific CTLs induced from colon cancer patients lysed the PAP+/HLA-A2+ colon cancer cell colo 205 specificly. HLA-A2-restricted PAP112-120 peptide could be used in the immunotherapy of colon cancer.
引文
[1]Fearon ER,Vogelstein B.A genetic model for colorectal tumorigenesis[J]. Cell,1990,61(5):759-767.
[2] Sidransky D.Tokino T.Hamilton S R.et al.Identification of ras oncogene mutations in the stool of patients with curable colorectal tumors[J].Science, l992,256(5053):l02-105.
[3]Smith-Ravin J,England J,Talbot IC,et al.Detection of c-Ki-ras mutations in faecal samples from sporadic colorectal cancer patients[J].Gut,1995, 36(1):81-86.
[4] Nallau P,Moser C,Weinland G.et al.Detection of K-ras mutations in stools of patients with colorectal cancer by mutant-enriched PCR[J].Int J Cancer, 1996, 66(3): 332-336.
[5] Jais P,Kapel N,Chosidow D.et al. Detection of protein p53 in the stool of patients with colorectal cancer[J].Gastroenterol Clin Biol,1997,21(10): 754-759.
[6] Minamoto T,Mai M,Ronai Z.K-ras mutation:early detection in molecular diagnosis and risk assessment of colorectal,pancreas and lung cancers[J], Cancer Detect Prev,2000,24(1):1-12.
[7]Orita M,Iwahana H, Kanazawa H,et al.Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms [J].Proc Natl Acad Sci USA,1989,86(8):2762-770.
[8]樊代明,牟震先,张学庸等.结肠癌单克隆MG3.5.7.10的制备及其相应抗原免疫组化定位研究[J].单克隆抗体通讯,1986,2(3):19-23.
[9]袁枚,刘成贵,李力等.抗结肠癌单克隆抗体CL-3,CL-4血清学诊断的应用[J].中国肛肺病杂志,1988,8(4):3-6.
[10] Symms M,Jager H R,Schmierer K,et a1.A review of structural magnetic resonance neuroimaging [J]. Neurol Neurosurg Psychiatry,2004,75(9):1235-1244.
[11] Gauvain K M,McKinstry R C,Mukherjee P,et a1.Evaluating pediatric brain tumor cellularity with diffusion-tensor imaging [J].AJR Am J Roentgenol, 2001, 177(2): 449-454.
[12]Rumboldt Z,Camacho DLA, Lake D, et al. Apparent diffusion coefficientsfor differentiation of cebellar tumors in childrenl[J],AJR Am J Neuroradiol, 2006, 27(6):1362-1369.
[13]Grossman S.Milos M L, Tekawa I S ,et a l. Colonoscopic screening of persons with suspected risk factors for colon cancer:ⅡPast history of colorectal neoplasms[J].Gastroenterology,1989,96(2):299-306.
[14]Yee J,Akerkar G A,Hung R K,et a1.Colorectal neoplasia:performance characteristics of colonography for detection in 300 patients[J].Radiology, 2001,219(3):685-692.
[15]Meier Ch,Wildermuth S,Feasibilty and potential of MR-colonography for evaluating colorectal cancer[J],Swiss Surg,2002,8(1):21-24.
[16]Seregni E,Coli A,Mazzucca N,et a1.Circulating tumor markers inbreastcancer[J].Eur J Nucl Med Mol Imaging,2004,31(suppl 1):Sl5—22.
[17]吴阶平,裘法祖,黄家驷.外科学[M].北京:人民卫生出版社,2004.1131.
[18]Chen CC,Yang SH.Is it resonable to add preoperative setum level of CEA and CA1 9-9 to staging for colorectal cacer[J].Surg Res,2005,124(2):169—174.
[19]Habermann JK,Roblick UJ,Luke BT,et a1.Increased serum levels of complement C3a anaphylatoxin indicate the presence of colorectal tumors[J]. Gastroenterology,2006,131(4):1020-1029.
[20]Celis J E,Kruhoffer M,Gromova I,et a1.Gene expression profiling: monitoring transcription and translation products using DNA microarrays and proteomics. [J].FEBS Lett,2000,480(1):2-16.
[21]Roessler M,Rollinger W,Palme s,et al.Identification of nicotinamide N-methyltransferase as a novel serum tumor marker for coloroctal cancer[J].Clin Cancer Res, 2005,11(18):6550-6557.
[22] Rayter Z,leicester R J, Mansi, J L.Adjuvant chemotherapy for colorectal cancer[J].Ann R Coll Surg Engl,1995,77(2):81-84.
[23] Beard S M,Holmes M,Price C,et al.Hepatic resection for colorectal liver metastases:A cost-effectiveness analysis[J].Ann Surg.2000;232(6):763-776.
[24]魏瑞,张阳德,何剪太等.三维适形放疗联合化疗治疗局部晚期和术后复发性直肠癌[J].现代肿瘤医学,2008,16(9):1584-1586.
[25]Agard C,Ligeza, C, Dupas, B, et al. Immune-dependent distant bystander effect after adenovirus-mediated suicide gene transfer in a rat model of liver colorectal metastasis[J]. Cancer Gene Ther,2001,8(2):128-136.
[26]Sivanandham M,Scoggin S D, Tanaka N, et al. Therapeutic effect of a vaccinia colon oncolysate prepared with interleukin-2-gene encoded vaccinia virus studied in a syngeneic CC-36 murine colon hepatic metastasis model[J].Cancer Immunol Immunother,1994,38(4): 259-264 .
[27]Chu Y,Hu H M,Winter H,et a1.Examining the immune response in sentinel lymph nodes of mice and men[J].Eur J Nucl Med ,1999,26(4): 50-53.
[28]姚德茂,王一理,马茂,等.自体肿瘤疫苗主动特异性免疫治疗进展期肿瘤的初步研究[J].中国免疫学杂志,2002,18(10):697-700.
[29]Broeke LT,Daschbach E,Thomas EK,et a1.Dendritic cell-induced activation of adaptive and innateantitumor immunity [J].Immunol,2003, 171(11):5842-5852.
[30]Eggert A A,Schreurs M W,Boerman OC,et al. Biodistribution and vaccine efficiency of murine dendritic cells are dependent on the route of administration[J]. Cancer Res,1999,59(14):3340-3345.
[31]Ribas A ,Bui LA ,Butterfield L H, et al. Antitumor protection using murine dendritic cells pulsed with acid-eluted peptides from in vivo grown tumors of different immunogencities[J]. Anticancer Res, 1999,19 (2A ):1652-1670.
[32]Uppenkamp M,Engert A,Diehl V,et al. Monoclonal antibody therapy with CAMPATH-1H in patients with relapsed high-and low-grade nonHodgkin's lymphomas: a multicenter phase I/II study[J]. Ann. Hematol, 2002,81(1):26-32.
[33]Fagerberg J,Hjelm AL,Ragnharnmar P,et al.Tumor regression in monoclonal antibody-treated patients correlates with the presente of anti- idiotypereactive T lymphocytes[J],Cancer Res,1995,55(9):1824-1827.
[34]Nakamura K,et al.Antibody Immunoconing Radiopharm.1992,5:359.
[35]刘艳艳,梁淑娟.白细胞介素-1在肿瘤靶向治疗策略中的价值.食品与药品[J].2009,11(3):53-57.
[36]施宗高,许良中,等.基因免疫抗肿瘤研究进展.癌症[J]. 2000, 19(6): 611-613.
[37]Whitesel J A, Donohue R E,Mani J H, et a1.Acid phosphatase:its influence on the managenment of carcinoma of the prostate[J].J Urol,1984, 131(2):70-72.
[38]Gutman AB, Gutman EB. An " acid " phosphatase occurring in the serum of patients with metastasizing carcinoma of the prostate gland[J]. J Clin Invest. 1938,17(4):473-478.
[39]Buamah P K.Johnson P.Skillen AW.Comparative study of the clinical usefulness of prostate specific antigen and prostatic acid phosphatase in prostatic disease[J].Br J Urol,1988,62(6):581-583.
[40]Arai Y,Yoshiki T,Okada K,et al .Multiple marker evaluation in prostatic cancer using prostatic specific antigen.gamma-seminoprotein and prostatic acid phosphatase[J]. Urol Int,1989,44(3):135-140.
[41]Solin T.Kontturi M,Pohlmann R,et a1.Gene expression and prostate specificity of human prostatic acid phosphatase(PAP):evaluation by RNA blot analyses[J].Biochim Biophys Acta,1990,1048(1):72-77.
[42]Nowels K,Kent E,Rinsho K,et a1.Prostate specific antigen and acid phophatase-reactive cells in cystitis cystica and glandularis[J].Arch Pathol Lab Med.1988,112(7):734-737.
[43]Epstein JI,Kuhajda F P,Lieberman P H.Prostate-specific acid phosphatase immunoreactivity in adenocarcinomas of the urinary bladder[J].Hum Patho1, 1986, 17(9):939-942.
[44]Quintero IB, Araujo CL, Pulkka AE, et al. Prostatic acid phosphatase is not a prostate specific target[J]. Cancer Res. 2007,67(14):6549-6554.
[45]Kaneko Y, Motoi N, Matsui A, et al. Neuroendocrine tumors of the liver and pancreas associated with elevated serum prostatic acid phosphatase[J]. Intern Med. 1995,34(9):886-891.
[46]王毅,王芳,宿晓云,等.前列腺酸性磷酸酶在非前列腺腺癌中的表达[J].第四军医大学学报.2007,28(19):1775-1777.
[47]Wang Y, Harada M, Yano H, et al. Prostatic acid phosphatase as a targetmolecule in specific immunotherapy for patients with non-prostate adenocarcinoma. Journal of Immunotherapy , 2005,28(6):535-541.
[48]Dattoli M ,Wallner K,True L,et a1.Long-term outcomes after treatment with external beam radiation therapy and palladium 103 for higher risk prostate carcinoma: influence of prostatic acid phosphatase[J].Cancer, 2003,97(4):979-983.
[49]Burch PA,Croghan GA,Gastineau DA,et a1.Immunotherapy(APC8015 Provenge) targeting prostatic acid phosphatase can induce durable remission of metastatic androgen-independent prostate cancer:a Phase 2 trial[J]. Prostate,2004, 60(3):197-204.
[50]Small E J,Fratesi P,Reese D M,et al. Immunotherapy of hormonerefractory prostate cancer with antigen-loaded dendritic cells [J].J Clin Oncol,2000, 18(23): 3894-3903.
[51] Seki K,Miyakoshi S,Lee GH,et a1.Prostatic acid phosphatase is a possible tumor marker for intravascular large B-cell lymphoma[J].Am J Surg Pathol, 2004, 28(10):1384-1388.
[52]Peshwa M V,Shi J D,Ruegg C,et al. Induction of prostate tumor-specific CD8+ cytotoxic T-lymphocytes in vitro using antigen-presenting cells pulsed with prostatic acid phosphatase peptide [J]. Prostate,1998,36(2):129-138.
[53]Kamoshida S,Tsutsumi Y.Extraprostatic localization of prostatic acid phosphatase and prostate-specific antigen : distribution in cloacogenic glandular epithelium and sex-dependent expression in human anal grand[J].Hum Pathol , 1990, 21(11) :1108-1111.
[54]Tepper SL,Jagirdar J,Heath D,et al. Homology between the female par -aurethral (Skene’s) glands and the prostate1. Immunohistochemical demodem- onstration[J]. Arch Pathol Lab Med,1984,108(5) :423-425.
[55]Mine T, Gouhara R, Hida N, et al. Immunological evaluation of CTL precursor-oriented vaccines for advanced lung cancer patients[J]. Cancer Sci., 2003,94: 548–556.
[56]Tanaka S, Harada M, Mine T, et al. Peptide vaccination for patients with melanoma and other types of cancer based on pre-existing peptide-specific ctotoxic T-lymphocyte precursors in the periphery [J] . Immunother, 2003, 26: 357–366.
[57]Noguchi M, Kobayashi K, Suetsugu N, et al. Induction of cellular and humoral immune responses to tumor cells and peptides in HLA-A24 positive hormone-refractory prostate cancer patients by peptide vaccination[J]. Prostate, 2003,57: 80–92.
[58]Sato Y, Shomura H, Maeda Y, et al. Immunological evaluation of peptide vaccination for patients with gastric cancer based on pre-existing cellular response to peptide[J]. Cancer Sci., 2003,94: 808–902.
[59]Noguchi M, Itoh K, Suekane S, et al. Phase I trial of patientoriented vaccination in HLA-A2 positive patients with metastatic hormone refractory prostate cancer[J]. Cancer Sci. 2004,95(1):77-84.
[60]DiFronzo LA, Gupta RK, Essner R, et al. Enhanced humoral immune response correlates with improved disease-free and overall survival in American Joint Committee on Cancer stage II melanoma patients receiving adjuvant polyvalent vaccine [J].Clin. Oncol,2002,20:3242–3248.
[61]Ohkouchi, S., Yamada, A., Imai, N., et al. Non-mutated tumor rejection antigen peptides elicit type-I allergy in the majority of healthy individuals[J]. Tissue Antigens,2002,59: 259–272.
[62]Kawamoto, N., Yamada, A., Ohkouchi, S., et al. IgG reactive to CTL-directed epitopes of self-antigens is either lacking or unbalanced in atopic dermatitis patients[J]. Tissue Antigens, 2003,61: 352–361.
[63]Robbins P F , Kawakami Y.Human tumor antigens recognized by T cells[J].Curr Opin Immunol,1996,8(5) :628-636.
[64]Harada M, Noguchi M, Itoh K.Target molecules in specific immunotherapy against prostate cancer[J]. Int J Clin Oncol , 2003 , 8(4) :193-199.
[2] Sidransky D.Tokino T.Hamilton S R.et al.Identification of ras oncogene mutations in the stool of patients with curable colorectal tumors[J].Science, l992,256(5053):l02-105.
[3]Smith-Ravin J,England J,Talbot IC,et al.Detection of c-Ki-ras mutations in faecal samples from sporadic colorectal cancer patients[J].Gut,1995, 36(1):81-86.
[4] Nallau P,Moser C,Weinland G.et al.Detection of K-ras mutations in stools of patients with colorectal cancer by mutant-enriched PCR[J].Int J Cancer, 1996, 66(3): 332-336.
[5] Jais P,Kapel N,Chosidow D.et al. Detection of protein p53 in the stool of patients with colorectal cancer[J].Gastroenterol Clin Biol,1997,21(10): 754-759.
[6] Minamoto T,Mai M,Ronai Z.K-ras mutation:early detection in molecular diagnosis and risk assessment of colorectal,pancreas and lung cancers[J], Cancer Detect Prev,2000,24(1):1-12.
[7]Orita M,Iwahana H, Kanazawa H,et al.Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms [J].Proc Natl Acad Sci USA,1989,86(8):2762-770.
[8]樊代明,牟震先,张学庸等.结肠癌单克隆MG3.5.7.10的制备及其相应抗原免疫组化定位研究[J].单克隆抗体通讯,1986,2(3):19-23.
[9]袁枚,刘成贵,李力等.抗结肠癌单克隆抗体CL-3,CL-4血清学诊断的应用[J].中国肛肺病杂志,1988,8(4):3-6.
[10] Symms M,Jager H R,Schmierer K,et a1.A review of structural magnetic resonance neuroimaging [J]. Neurol Neurosurg Psychiatry,2004,75(9):1235-1244.
[11] Gauvain K M,McKinstry R C,Mukherjee P,et a1.Evaluating pediatric brain tumor cellularity with diffusion-tensor imaging [J].AJR Am J Roentgenol, 2001, 177(2): 449-454.
[12]Rumboldt Z,Camacho DLA, Lake D, et al. Apparent diffusion coefficientsfor differentiation of cebellar tumors in childrenl[J],AJR Am J Neuroradiol, 2006, 27(6):1362-1369.
[13]Grossman S.Milos M L, Tekawa I S ,et a l. Colonoscopic screening of persons with suspected risk factors for colon cancer:ⅡPast history of colorectal neoplasms[J].Gastroenterology,1989,96(2):299-306.
[14]Yee J,Akerkar G A,Hung R K,et a1.Colorectal neoplasia:performance characteristics of colonography for detection in 300 patients[J].Radiology, 2001,219(3):685-692.
[15]Meier Ch,Wildermuth S,Feasibilty and potential of MR-colonography for evaluating colorectal cancer[J],Swiss Surg,2002,8(1):21-24.
[16]Seregni E,Coli A,Mazzucca N,et a1.Circulating tumor markers inbreastcancer[J].Eur J Nucl Med Mol Imaging,2004,31(suppl 1):Sl5—22.
[17]吴阶平,裘法祖,黄家驷.外科学[M].北京:人民卫生出版社,2004.1131.
[18]Chen CC,Yang SH.Is it resonable to add preoperative setum level of CEA and CA1 9-9 to staging for colorectal cacer[J].Surg Res,2005,124(2):169—174.
[19]Habermann JK,Roblick UJ,Luke BT,et a1.Increased serum levels of complement C3a anaphylatoxin indicate the presence of colorectal tumors[J]. Gastroenterology,2006,131(4):1020-1029.
[20]Celis J E,Kruhoffer M,Gromova I,et a1.Gene expression profiling: monitoring transcription and translation products using DNA microarrays and proteomics. [J].FEBS Lett,2000,480(1):2-16.
[21]Roessler M,Rollinger W,Palme s,et al.Identification of nicotinamide N-methyltransferase as a novel serum tumor marker for coloroctal cancer[J].Clin Cancer Res, 2005,11(18):6550-6557.
[22] Rayter Z,leicester R J, Mansi, J L.Adjuvant chemotherapy for colorectal cancer[J].Ann R Coll Surg Engl,1995,77(2):81-84.
[23] Beard S M,Holmes M,Price C,et al.Hepatic resection for colorectal liver metastases:A cost-effectiveness analysis[J].Ann Surg.2000;232(6):763-776.
[24]魏瑞,张阳德,何剪太等.三维适形放疗联合化疗治疗局部晚期和术后复发性直肠癌[J].现代肿瘤医学,2008,16(9):1584-1586.
[25]Agard C,Ligeza, C, Dupas, B, et al. Immune-dependent distant bystander effect after adenovirus-mediated suicide gene transfer in a rat model of liver colorectal metastasis[J]. Cancer Gene Ther,2001,8(2):128-136.
[26]Sivanandham M,Scoggin S D, Tanaka N, et al. Therapeutic effect of a vaccinia colon oncolysate prepared with interleukin-2-gene encoded vaccinia virus studied in a syngeneic CC-36 murine colon hepatic metastasis model[J].Cancer Immunol Immunother,1994,38(4): 259-264 .
[27]Chu Y,Hu H M,Winter H,et a1.Examining the immune response in sentinel lymph nodes of mice and men[J].Eur J Nucl Med ,1999,26(4): 50-53.
[28]姚德茂,王一理,马茂,等.自体肿瘤疫苗主动特异性免疫治疗进展期肿瘤的初步研究[J].中国免疫学杂志,2002,18(10):697-700.
[29]Broeke LT,Daschbach E,Thomas EK,et a1.Dendritic cell-induced activation of adaptive and innateantitumor immunity [J].Immunol,2003, 171(11):5842-5852.
[30]Eggert A A,Schreurs M W,Boerman OC,et al. Biodistribution and vaccine efficiency of murine dendritic cells are dependent on the route of administration[J]. Cancer Res,1999,59(14):3340-3345.
[31]Ribas A ,Bui LA ,Butterfield L H, et al. Antitumor protection using murine dendritic cells pulsed with acid-eluted peptides from in vivo grown tumors of different immunogencities[J]. Anticancer Res, 1999,19 (2A ):1652-1670.
[32]Uppenkamp M,Engert A,Diehl V,et al. Monoclonal antibody therapy with CAMPATH-1H in patients with relapsed high-and low-grade nonHodgkin's lymphomas: a multicenter phase I/II study[J]. Ann. Hematol, 2002,81(1):26-32.
[33]Fagerberg J,Hjelm AL,Ragnharnmar P,et al.Tumor regression in monoclonal antibody-treated patients correlates with the presente of anti- idiotypereactive T lymphocytes[J],Cancer Res,1995,55(9):1824-1827.
[34]Nakamura K,et al.Antibody Immunoconing Radiopharm.1992,5:359.
[35]刘艳艳,梁淑娟.白细胞介素-1在肿瘤靶向治疗策略中的价值.食品与药品[J].2009,11(3):53-57.
[36]施宗高,许良中,等.基因免疫抗肿瘤研究进展.癌症[J]. 2000, 19(6): 611-613.
[37]Whitesel J A, Donohue R E,Mani J H, et a1.Acid phosphatase:its influence on the managenment of carcinoma of the prostate[J].J Urol,1984, 131(2):70-72.
[38]Gutman AB, Gutman EB. An " acid " phosphatase occurring in the serum of patients with metastasizing carcinoma of the prostate gland[J]. J Clin Invest. 1938,17(4):473-478.
[39]Buamah P K.Johnson P.Skillen AW.Comparative study of the clinical usefulness of prostate specific antigen and prostatic acid phosphatase in prostatic disease[J].Br J Urol,1988,62(6):581-583.
[40]Arai Y,Yoshiki T,Okada K,et al .Multiple marker evaluation in prostatic cancer using prostatic specific antigen.gamma-seminoprotein and prostatic acid phosphatase[J]. Urol Int,1989,44(3):135-140.
[41]Solin T.Kontturi M,Pohlmann R,et a1.Gene expression and prostate specificity of human prostatic acid phosphatase(PAP):evaluation by RNA blot analyses[J].Biochim Biophys Acta,1990,1048(1):72-77.
[42]Nowels K,Kent E,Rinsho K,et a1.Prostate specific antigen and acid phophatase-reactive cells in cystitis cystica and glandularis[J].Arch Pathol Lab Med.1988,112(7):734-737.
[43]Epstein JI,Kuhajda F P,Lieberman P H.Prostate-specific acid phosphatase immunoreactivity in adenocarcinomas of the urinary bladder[J].Hum Patho1, 1986, 17(9):939-942.
[44]Quintero IB, Araujo CL, Pulkka AE, et al. Prostatic acid phosphatase is not a prostate specific target[J]. Cancer Res. 2007,67(14):6549-6554.
[45]Kaneko Y, Motoi N, Matsui A, et al. Neuroendocrine tumors of the liver and pancreas associated with elevated serum prostatic acid phosphatase[J]. Intern Med. 1995,34(9):886-891.
[46]王毅,王芳,宿晓云,等.前列腺酸性磷酸酶在非前列腺腺癌中的表达[J].第四军医大学学报.2007,28(19):1775-1777.
[47]Wang Y, Harada M, Yano H, et al. Prostatic acid phosphatase as a targetmolecule in specific immunotherapy for patients with non-prostate adenocarcinoma. Journal of Immunotherapy , 2005,28(6):535-541.
[48]Dattoli M ,Wallner K,True L,et a1.Long-term outcomes after treatment with external beam radiation therapy and palladium 103 for higher risk prostate carcinoma: influence of prostatic acid phosphatase[J].Cancer, 2003,97(4):979-983.
[49]Burch PA,Croghan GA,Gastineau DA,et a1.Immunotherapy(APC8015 Provenge) targeting prostatic acid phosphatase can induce durable remission of metastatic androgen-independent prostate cancer:a Phase 2 trial[J]. Prostate,2004, 60(3):197-204.
[50]Small E J,Fratesi P,Reese D M,et al. Immunotherapy of hormonerefractory prostate cancer with antigen-loaded dendritic cells [J].J Clin Oncol,2000, 18(23): 3894-3903.
[51] Seki K,Miyakoshi S,Lee GH,et a1.Prostatic acid phosphatase is a possible tumor marker for intravascular large B-cell lymphoma[J].Am J Surg Pathol, 2004, 28(10):1384-1388.
[52]Peshwa M V,Shi J D,Ruegg C,et al. Induction of prostate tumor-specific CD8+ cytotoxic T-lymphocytes in vitro using antigen-presenting cells pulsed with prostatic acid phosphatase peptide [J]. Prostate,1998,36(2):129-138.
[53]Kamoshida S,Tsutsumi Y.Extraprostatic localization of prostatic acid phosphatase and prostate-specific antigen : distribution in cloacogenic glandular epithelium and sex-dependent expression in human anal grand[J].Hum Pathol , 1990, 21(11) :1108-1111.
[54]Tepper SL,Jagirdar J,Heath D,et al. Homology between the female par -aurethral (Skene’s) glands and the prostate1. Immunohistochemical demodem- onstration[J]. Arch Pathol Lab Med,1984,108(5) :423-425.
[55]Mine T, Gouhara R, Hida N, et al. Immunological evaluation of CTL precursor-oriented vaccines for advanced lung cancer patients[J]. Cancer Sci., 2003,94: 548–556.
[56]Tanaka S, Harada M, Mine T, et al. Peptide vaccination for patients with melanoma and other types of cancer based on pre-existing peptide-specific ctotoxic T-lymphocyte precursors in the periphery [J] . Immunother, 2003, 26: 357–366.
[57]Noguchi M, Kobayashi K, Suetsugu N, et al. Induction of cellular and humoral immune responses to tumor cells and peptides in HLA-A24 positive hormone-refractory prostate cancer patients by peptide vaccination[J]. Prostate, 2003,57: 80–92.
[58]Sato Y, Shomura H, Maeda Y, et al. Immunological evaluation of peptide vaccination for patients with gastric cancer based on pre-existing cellular response to peptide[J]. Cancer Sci., 2003,94: 808–902.
[59]Noguchi M, Itoh K, Suekane S, et al. Phase I trial of patientoriented vaccination in HLA-A2 positive patients with metastatic hormone refractory prostate cancer[J]. Cancer Sci. 2004,95(1):77-84.
[60]DiFronzo LA, Gupta RK, Essner R, et al. Enhanced humoral immune response correlates with improved disease-free and overall survival in American Joint Committee on Cancer stage II melanoma patients receiving adjuvant polyvalent vaccine [J].Clin. Oncol,2002,20:3242–3248.
[61]Ohkouchi, S., Yamada, A., Imai, N., et al. Non-mutated tumor rejection antigen peptides elicit type-I allergy in the majority of healthy individuals[J]. Tissue Antigens,2002,59: 259–272.
[62]Kawamoto, N., Yamada, A., Ohkouchi, S., et al. IgG reactive to CTL-directed epitopes of self-antigens is either lacking or unbalanced in atopic dermatitis patients[J]. Tissue Antigens, 2003,61: 352–361.
[63]Robbins P F , Kawakami Y.Human tumor antigens recognized by T cells[J].Curr Opin Immunol,1996,8(5) :628-636.
[64]Harada M, Noguchi M, Itoh K.Target molecules in specific immunotherapy against prostate cancer[J]. Int J Clin Oncol , 2003 , 8(4) :193-199.