I HC QUC GIA H NI TRNG I HC KHOA HC T NHIấN - Lấ LAN PHNG XC NH KHNG NGUYấN PHN NG MIN DCH CA BNH NHN UNG TH GAN LUN VN THC S KHOA HC H Ni - 2012 I HC QUC GIA H NI TRNG I HC KHOA HC T NHIấN - Lờ Lan Phng XC NH KHNG NGUYấN PHN NG MIN DCH CA BNH NHN UNG TH GAN Chuyờn ngnh: Sinh hc thc nghim Mó s: CH.60.42.30 LUN VN THC S KHOA HC NGI HNG DN KHOA HC: PGS.TS Trnh Hng Thỏi H Ni - 2012 LI CM N Tụi xin by t lũng bit n sõu sc ti PGS TS Trnh Hng Thỏi, ngi thy ó tn tỡnh ch bo, hng dn tụi sut quỏ trỡnh hc tp, nghiờn cu khoa hc v thc hin lun ny Tụi cng xin trõn trng cm n cỏc thy giỏo, cụ giỏo Khoa Sinh hc, Trng i hc Khoa hc T nhiờn ó tn tỡnh ging dy, dỡu dt tụi thi gian hc ti Trng Tụi xin c gi li cm n chõn thnh ti cỏc anh, ch hc viờn v cỏc bn sinh viờn lm vic ti Phũng Proteomics v Sinh hc cu trỳc thuc Phũng thớ nghim Trng im Cụng ngh Enzyme v Protein, Trng i hc Khoa hc T nhiờn ó tn tỡnh ch bo v giỳp tụi sut quỏ trỡnh hc v thc hin lun ti Phũng Trong quỏ trỡnh thc hin lun vn, tụi ó nhn c rt nhiu s quan tõm giỳp ca cỏc cỏn b, nhõn viờn thuc Khoa T bo - Gii phu bnh, Bnh vin K2, Tam Hip, H Ni Tụi xin chõn thnh cm n Cui cựng, tụi vụ cựng bit n gia ỡnh v bn bố ó khớch l, ng viờn, giỳp tụi sut thi gian qua Lun c thc hin vi s h tr kinh phớ t ti mó s KLEPT-09-02 H Ni, thỏng nm 2012 Hc viờn Lờ Lan Phng MC LC BNG Kí HIU V CC CH VIT TT a DANH MC CC BNG d DANH MC CC HèNH e M U .1 Chng - TNG QUAN 1.1 TNG QUAN V UNG TH GAN .3 1.1.1 Phõn loi ung th gan nguyờn phỏt 1.1.2 Nguyờn nhõn dn ti ung th gan .4 1.1.3 Cỏc giai on ca ung th gan 1.1.4 Cỏc phng phỏp chn oỏn ung th gan 1.2 NGHIấN CU CH TH SINH HC I VI UNG TH 1.2.1 Ch th sinh hc i vi ung th 1.2.2 Tip cn nghiờn cu ch th sinh hc i vi ung th .10 1.2.3 Cỏc loi ch th sinh hc i vi ung th gan 11 1.3 MIN DCH CHNG UNG TH 13 1.3.1 ỏp ng dch chng ung th 13 1.3.2 Khỏng nguyờn ung th 13 1.3.3 Ch th ung th liờn quan n ỏp ng dch 14 1.4 NGHIấN CU PROTEOMICS MIN DCH UNG TH GAN 15 1.4.1 Khỏi quỏt v proteomics v ng dng ca proteomics 15 1.4.2 H protein gan ngi .16 1.4.3 Nghiờn cu proteomics dch ung th gan 17 Chng - NGUYấN LIU V PHNG PHP 24 2.1 NGUYấN LIU .24 2.1.1 i tng nghiờn cu .24 2.1.2 Húa cht 24 2.1.3 Thit b 25 2.2 PHNG PHP NGHIấN CU 25 2.2.1 X lý mu mụ gan 25 2.2.2 nh lng protein theo phng phỏp Bradford 27 2.2.3 in di hai chiu .27 2.2.4 Western Blot 28 2.2.5 Phõn tớch hỡnh nh bn gel in di hai chiu, kt qu Western Blot 29 2.2.6 Ct v thy phõn spot trờn bn gel in di hai chiu 30 2.2.7 Phõn tớch ph v nhn dng protein 31 Chng - KT QU V BN LUN 33 3.1 TCH CHIT PROTEIN T Mễ GAN 33 3.2 PHN TCH HèNH NH BN GEL IN DI HAI CHIU 34 3.2.1 Phõn tỏch h protein mụ gan trờn bn gel din di hai chiu 34 3.2.2 Phõn tớch biu hin ca protein trờn bn gel in di hai chiu .36 3.3 NHN DNG PROTEIN BNG PHNG PHP MALDI-TOF MS 41 3.4 THM TCH MIN DCH - WESTERN BLOT HAI CHIU 44 3.4.1 Xỏc nh t l pha loóng khỏng th bc mt thớch hp 44 3.4.2 Kt qu Western Blot hai chiu 46 3.5 C IM, VAI TRề CA CC PROTEIN BIU HIN KHC BIT 47 3.5.1 Protein liờn quan n chu trỡnh t bo v apoptosis 50 3.5.2 Protein liờn quan n quỏ trỡnh dch v bo v c th 52 3.5.3 Protein tham gia cu trỳc t bo 53 3.5.4 Protein tham gia cỏc quỏ trỡnh trao i cht 54 3.5.5 Protein liờn quan n quỏ trỡnh phiờn mó, dch mó v ci bin 55 3.5.6 Protein cú phn ng dch vi huyt tng .56 KT LUN .60 KIN NGH 61 TI LIU THAM KHO 62 PH LC i Ph lc Danh sỏch bnh nhõn ung th t bo gan i Ph lc Phõn tỏch protein mụ gan ca bnh nhõn HCC trờn bn gel in di hai chiu ii Ph lc Kt qu nhn dng protein bng c s d liu NCBI s dng phn mm Mascot vi Ph lc Kt qu phn ng Western Blot mu mụ gan ca bnh nhõn HCC viii BNG Kí HIU V CC CH VIT TT 1AT Alpha-1-antitrypsin ABC Ammonium bicarbonate AcCN Acetonitrile ACTB Beta-actin AFP Alpha fetoprotein AFB1 Aflatoxin B1 CATD Cathepsin D CHAPS 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonic acid CHCA -Cyano-4-hydroxycinnamic acid cs Cng s Da Dalton DTT Dithiothreitol ELISA Enzyme-linked immunosorbent assay (Xột nghim hp ph dch liờn kt enzyme) GS Glutamine synthetase HBV Hepatitis B virus (Virus viờm gan B) HBsAg Hepatitis B surface Antigen (Khỏng nguyờn b mt virus viờm gan B) HCC Hepatocellular carcinoma (Ung th t bo biu mụ gan) HCV Hepatitis C virus (Virus viờm gan C) HLPP Human Liver Proteome Project (D ỏn Proteome gan ngi) HPT Haptoglobin HUPO Human Proteome Organisation (T chc Proteome ngi) HRP Horse - radish peroxidase a HSP Heat shock protein (Protein sc nhit) IAA Iodoacetamide IEF Isoelectric Focusing (in di phõn vựng ng in) IPG Immobilized pH gradient (Gradient pH c nh) LC-MS/MS Liquid Chromatography coupled with tandem Mass Spectrometry (Sc ký lng kt ni vi ph) MALDI-TOF Matrix-assisted laser desorption ionization time-of-flight MFAP3 Microfibrillar-associated protein MHC Mayjor Histocampatibility Complex (Phc h phự hp t chc mụ ch yu) MS Mass spectrometry (Khi ph) Mw Molecular weight (Khi lng phõn t) PBS Phosphate Buffer Saline (m mui phosphate) PCNA Proliferating cell nuclear antigen PDIA1 Protein disulfide-isomerase PDIA3 Protein disulfide-isomerase A3 PMF Peptide Mass Fingerprint (c trng peptide) PMSF Phenylmethanesulfonyl fluoride PVDF Polyvinylidene fluoride p53 Cellular tumor antigen p53 SDS Sodium dodecyl sulfate SDS-PAGE SDS-Polyacrylamide Gel Electrophoresis (in di trờn gel polyacrylamide cú SDS) SEREX Serological Analysis of Recombinant cDNA Expression Libraries (Phõn tớch huyt bng th vin biu hin cDNA tỏi t hp) SODC Superoxide dismutase [Cu-Zn] b SODM Superoxide dismutase [Mn] Mn-SOD Manganese superoxide dismutase Spot im protein ST7L Suppressor of tumorigenicity protein-like TAA Tumour Associated Antigen (Khỏng nguyờn liờn quan n ung th) TAP Transporters of Antigen Peptide (Phõn t chuyn peptide khỏng nguyờn) TFA Trifluoroacetic acid TNM Tumor Node Metastasis (Kớch thc u - Hch Lympho - Di cn) TSA Tissue Specific Antigen (Khỏng nguyờn c hiu mụ) TSTA Tumour Specific Transplantation Antigen (Khỏng nguyờn ghộp c hiu ung th) c DANH MC CC BNG Bng Tiờu chun phõn k lõm sng bnh ung th gan Bng Cỏc ch th ung th gan mi c cụng b (20 nm gn õy) 12 Bng Cỏc húa cht chớnh s dng nghiờn cu 24 Bng Cỏc bc chy in di ng in trờn strip di 7cm v 17cm 28 Bng Tng s spot trờn mi bn gel in di hai chiu 36 Bng Thng kờ cỏc spot biu hin khỏc bit gia mụ gan ung th so vi mụ gan bỡnh thng ca tng bnh nhõn trờn bn gel 2-DE 17cm, pH - 10 39 Bng Thng kờ cỏc spot biu hin khỏc bit gia mụ gan ung th so vi mụ gan bỡnh thng ca tng bnh nhõn trờn bn gel 2-DE 7cm, pH - 39 Bng Danh sỏch cỏc protein biu hin khỏc bit trờn bn gel ca mụ gan ung th so vi mụ gan bỡnh thng c xỏc nh da trờn c s d liu EMBL 40 Bng Danh sỏch cỏc protein c xỏc nh bng MALDI-TOF MS t bn gel mụ gan ung th so sỏnh vi mụ gan i chng ca bnh nhõn HCC .42 Bng 10 Túm tt chc nng chớnh ca cỏc protein biu hin khỏc bit gia mụ gan ung th so vi mụ gan i chng ó c nhn dng 48 d Luận văn cao học Lê Lan Ph-ơng KT LUN Bc u phõn tớch proteomics dch ca bnh nhõn ung th gan, chỳng tụi rỳt mt s kt lun sau: Bng k thut in di hai chiu ó phõn tỏch c cỏc protein mụ gan ung th v mụ gan bỡnh thng ca bnh nhõn ung th gan thnh cỏc spot riờng r trờn cỏc bn gel in di ó xỏc nh c cỏc spot protein biu hin khỏc bit gia mụ gan ung th v mụ gan bỡnh thng ca tng bnh nhõn ung th gan Trong ú cú 52 spot khỏc bit chung cỏc bnh nhõn gm: 23 spot biu hin tng, 18 spot biu hin gim, spot ch xut hin bn gel mụ ung th, spot khụng xut hin bn gel mụ ung th ó nhn dng c 52 spot tng ng vi 38 protein ó c nh danh v 12 protein gi thuyt Cỏc protein c chia thnh nhúm chc nng, bao gm: nhúm protein liờn quan n chu trỡnh t bo v apoptosis; protein liờn quan n quỏ trỡnh dch v bo v c th; protein tham gia cu trỳc t bo; protein tham gia cỏc quỏ trỡnh trao i cht v protein tham gia vo quỏ trỡnh phiờn mó, dch mó v ci bin sau phiờn mó, dch mó Trong ú, protein tham gia vo quỏ trỡnh dch v bo v c th chim t l cao nht (~29%), thp nht l nhúm protein tham gia vo cu trỳc t bo (~5%) S dng kit ECL Advance Western Blotting Detection , ó xac inh c t l pha loóng tụ i u khỏng th bc mụ t v th m tỏch dch thnh cụng trờn cp bn gel in di chiu ca bnh nhõn ung th gan Bng k thut thm tỏch dch Western blot hai chiu ó xỏc nh c protein cú phn ng dch la : protein ST7L biu hin tng v protein MHC lp I, HSP27, aldehyde dehydrogenase biu hin gim mụ gan ung th so vi mụ gan bỡnh thng 60 Luận văn cao học Lê Lan Ph-ơng KIN NGH Tri qua nghiờn cu thc t, chỳng tụi a mt s kin ngh sau: Tip tc nhn dng cỏc spot protein biu hin khỏc bit gia mụ ung th so vi mụ gan thng v kt hp vi kt qu phõn tớch thm tỏch dch Western blot hai chiu trờn s lng bnh nhõn ln hn nhm tỡm cỏc protein cú tớnh khỏng nguyờn c trng cho ung th gan Tip tc thu thp mu v phõn tớch proteomics dch mụ gan ca bnh nhõn ung th gan cỏc giai on ung th khỏc nhm phõn tớch s bin i thnh phn cỏc protein liờn quan n bnh Thu thp mu huyt tng ca bnh nhõn viờm gan B v C ang tin trin thnh ung th gan v bnh nhõn ung th gan cú tin s nhim HBV v HCV nhm tỡm hiu mi liờn h gia viờm gan B , C v ung th gan trờn cac ụ i t ng bờ nh nhõn ung th gan Viờ t Nam 61 Luận văn cao học Lê Lan Ph-ơng TI LIU THAM KHO Ting Vit Phan Vn Chi (2006), PROTEOMICS Khoa hc v h protein, Vin Khoa hc v Cụng ngh Vit Nam, H Ni Phan Vn Chi (2009), Thc trng nghiờn cu v Proteomics, Hi tho VNProteomics ln 1, Nh xut bn Khoa hc t nhiờn v cụng ngh Ngc Liờn (2004), Min dch hc c s, Nh xut bn i hc Quc gia H Ni Hong Vn Sn (2009), Proteomics lõm sng, Hi tho VNProteomics ln 1, Nh xut bn Khoa hc t nhiờn v cụng ngh Hong Vn Sn (2011), Cỏc ch t ung th gan, Tp y hc Vit Nam, 348, 5-11 Bựi Phng Tho, Lờ Lan Phng, Trnh Hng Thỏi (2011), Phõn tớch cỏc protein biu hin khỏc bit ca mụ gan bnh nhõn ung th gan, Tp y hc Vit Nam, 348, 67-71 Trng i hc Y H Ni (2006), Min dch hc, Nh xut bn Y hc, H Ni Ting Anh American Cancer Society (2012), Cancer Facts & Figures, National Health Council Bradford M M (1976), A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Analytical Biochemistry, 72, 248-254 10 Chen Y (2007), Identification of tumor-associated antigens as markers for immunodiagnosis of human cancers, ETD Collection for University of Texas, El Paso, Paper AAI3262904 62 Luận văn cao học Lê Lan Ph-ơng 11 Chen Y., Zhou Y., Qiu S., Wang K., Liu S., Peng X.X., Li J., Tan E.M., Zhang J.Y (2010), Autoantibodies to tumor-associated antigens combined with abnormal alpha-fetoprotein enhance immunodiagnosis of hepatocellular carcinoma, Cancer Letters, 289(1), 32-39 12 Diamandis E.P (2004), Identification of serum amyloid A protein as a potentially useful biomarker for nasopharyngeal carcinoma, Clinical Cancer Research, 10, 5293-5294 13 Farazi P.A., Depinho R.A (2006), Hepatocellular carcinoma pathogenses: from genes to evironment, Nature Reviews Cancer, 6, 674-687 14 Farombi E.O (2006), Aflatoxin contamination of foods in developing countries: Implications for hepatocellular carcinoma and chemopreventive strategies, African Journal of Biotechnology, 5(1), 001-014 15 Govekar R and Zingde S.M (2007), Cancer Proteomics: How far are we from the Clinics? , International Journal of Human Genetics, 7(1), 91-97 16 He Q.Y., Lau G.K.K., Zhou Y., Yuen S.T., Lin M.C., Kung H.F and Chiu J.F (2003), Serum biomarkers of hepatitis B virus infected liver inflammation: A proteomic study, Proteomics, 3(5), 666-674 17 Jain K.K (2002), Role of proteomics in diagnosis of cancer, Technology in Cancer Research and Treatment, 1(4), 281-286 18 Jemal A., Bray F., Center M M., Ferlay J., Ward E., Forman D., (2011), Global Cancer Statistics, CA Cancer J Clin, 61, 69-90 19 Kim W., Lim S.O., Kim J.S., Ryu Y.H., Byeon J.Y., Kim H.J., Kim Y.L., Heo J.S., Park Y.M., and Jung G (2003), Comparison of Proteome between Hepatitis B virus-and Hepatitis C virus-Associated Carcinoma, Clinical Cancer Research, 9, 5493-5500 63 Hepatocellular Luận văn cao học Lê Lan Ph-ơng 20 Le L.P., Pham A.T., Trinh H.T (2010), Proteomics analysis of human hepatocellular carcinoma, Vietnam National University Journal of Science, Natural Science and Technology, 26(1), 85-90 21 Li C., Tan Y.X., Zhou H., Ding S.J., Li S.J., Ma D.J., Man X.B., Hong Y., Zhang L., Li L., Xia Q.C., Wu J.R., Wang H.Y., Zeng R (2005), Proteomic analysis of hepatitis B virus-associated hepatocellular carcinoma: Identification of potential tumor markers, Proteomics, 5(4), 1125-1139 22 Li L., Chen S.H., Yu C.H., Li Y.M and Wang S.Q (2008), Identification of Hepatocellular-Carcinoma-Associated Antigens and Autoantibodies by Serological Proteome Analysis Combined with Protein Microarray, Journal of Proteome Research, 7(2), 611-620 23 Liebler D.C (2002), Introduction to proteomics, Human Press, Totowa, New Jersey 24 Liu W., Peng B., Lu Y., Xu W., Qian W., Zhang J.Y (2011), Autoantibodies to tumor-associated antigens as biomarkers in cancer immunodiagnosis, Autoimmunity Reviews, 10(6), 331-335 25 Long J., Lang Z.W., Wang H.G., Wang T.L., Wang B.E (2010), Glutamine synthetase as an early marker for hepatocellular carcinoma based on proteomic analysis of resected small hepatocellular carcinomas, Hepatobiliary Pacreat Diseases International, 9(3), 296-305 26 Looi K.S., Nakayasu E.S., Diaz R.A., Tan E.M., Almeida I.C., Zhang J.Y (2008), Using Proteomic Approach to Identify Tumor-Associated Antigens as Markers in Hepatocellular Carcinoma, Journal of Proteome Research, 7(9), 4004-4012 27 Lu H., Goodell V., Disis M.L (2008), Humoral immunity directed against tumor-associated antigens as potential biomarkers for the early diagnosis of cancer, Journal of Proteome Research, 7(4), 1388-1394 64 Luận văn cao học Lê Lan Ph-ơng 28 Martin K., Ricciardelli C., Hoffmann P., K.Oehker M., (2011), Exploring the Immunoproteome for Ovarian Cancer Biomarker Discovery, International Journal of Molecular Sciences, 12, 410-28 29 Mendy M., Walton R (2009), Molecular pathogenesis and early detection of Hepatocellular perspectives from West Africa, Cancer Letters; 286(1), 44-51 30 Polanski M., Anderson N.L (2006), A list of candidate cancer biomarkers for targeted proteomics, Biomark Insights, 1, 1-48 31 Qin L.X., Tang Z.Y (2002), The prognostic molecular markers in hepatocellular carcinoma, World Journal of Gastroenterology, 8(3), 385-392 32 Rubin P., Hansen J.T (2008), TNM Staging Atlas, Lippincott Williams & Wilkins, a Wolters Kluwer Business, Bogota, Colombia 33 Takashima M., Kuramitsu Y., Yokoyama Y., Iizuka N., Harada T., Fujimoto M., Sakaida I., Okita K., Oka M., (2006), Proteomic analysis of autonantibides in patients with hepatocellular carcinoma, Proteomics, 6(13), 3894-3900 34 Tan H., Low J., Lim S.G., Chung M.C (2009), Serum autoantibodies as biomarkers for early cancer detection, FEBS journal, 276(23), 6880-6904 35 Tjalsma H., Schaeps R.M., Swinkels D.W (2008), Immunoproteomics: From biomarker discovery to diagnostic applications, Proteomics Clinical Appllications, 2(2), 167-180 36 Tran T.T., Le L.P, Trinh H.T (2011), Analysis of differentially expressed proteins of plasma in hepatocellular carcinoma, Vietnam National University Journal of Science, Natural Science and Technology, 27(2), 279-284 37 Westermeier R., Naven T (2002), Proteomics in Practice, Wiley-VCH Verlag-GmbH, Freiburg 65 Luận văn cao học Lê Lan Ph-ơng 38 Wong C.H., Chan S.K.P., Chan H.L.Y., Tsui S.K.W (2006), The Molecular Diagnosis of Hepatitis B Virus-Associated Hepatocellular Carcinoma, Clinical Laboratory Sciences, 43(1), 69-101 39 Yao D.F., Dong Z.Z., Yao M (2007), Specific molecular markers in hepatocellular carcinoma, Hepatobiliary Pancreat Dis Int, 6(3), 241-247 40 Zhang J.Y., Megliorino R., Peng X.X., Tan E.M., Chen Y., Chan E.K (2007), Antibody detection using tumor-associated antigen mini-array in immunodiagnosing human hepatocellular carcinoma, Journal of Hepatology, 46(1), 107-114 Cụng c World Wide Web 41 http://benhvienk.com/index.php/vi/pcut/tin-tuc-phong-chong-ung-thu/677dau-hieu-cua-benh-ung-thu-gan (5/6/2012) 42 http://cancerhelp.cancerresearchuk.org/type/liver-cancer/about/types-ofprimary-liver-cancer (14/02/2012) 43 http://expasy.org/swiss-2dpage/viewer (5/6/2012) 44 http://igrid-ext.cryst.bbk.ac.uk/WWW/PhD_thesis.htm (10/06/2012) 45 http://www.cancer.gov/cancertopics/pdq/treatment/adult-primaryliver/HealthProfessional/page3 (14/02/2012) 46 http://www.cancer.gov/dictionary?cdrid=45618 (15/03/2012) 47 http://www.hupo.org/research/hlpp/ (25/04/2012) 48 http://www.ncbi.nlm.nih.gov/gene/54879 (10/06/2012) 49 http://www.mayoclinic.com/health/medical/IM04073 (30/03/2012) 50 http://www.medicinenet.com/liver_biopsy/article.htm (15/03/2012) 51 http://www.uniprot.org/ (05/06/2012) 52 www.matrixscience.com (05/06/2012) 66 PH LC Ph lc Danh sỏch bnh nhõn ung th t bo gan STT H v tờn Gii tớnh Tui Mó Giai on Ngy ly mu Bựi Xuõn Nam 61 8261 T2N0M0 05/11/2010 Nguyn Th M N 69 8327 T2N0M0 10/11/2010 Phm Vn T Nam 62 8344 T1N0M0 Phm ng N Nam 54 8460 T2N0M0 18/11/2010 Nguyn Vn B Nam 49 8754 T2N0M0 10/12/2010 Trn Vn S Nam 49 8786 T1N0M0 12/12/2010 Nguyn Trng P Nam 56 8977 T2N0M0 Phm Vn Nam 64 8935 T2N0M0 Lc Th T N 63 8996 T3N1M0 10 Nguyn Vn C Nam 59 9242 T2N0M0 11 V Vn M Nam 65 9543 T2N0M0 24/12/2010 21/01/2011 24/02/2011 12 Nguyn Th T N 63 9567 T1N0M0 13 Trn Hi A Nam 46 10480 T2N0M0 04/05/2011 14 Nguyn Ngc T Nam 51 13282 T2N0M0 15/11/2011 15 Triu Vn L Nam 41 13994 T1N0M0 06/01/2012 i Ph lc Phõn tỏch protein mụ gan ca bnh nhõn HCC trờn bn gel in di hai chiu Chiu 1: in di ng in s dng IPG strip di 17cm, pH - 10 (A1 v B1); IPG strip di 7cm, pH - (A2 v B2); Chiu 2: in di SDS-PAGE trờn gel polyacrylamide 10% A - Mụ gan bỡnh thng; B - Mụ gan ung th - spot protein ST7L; - spot protein aldehyde dehydrogenase; - spot protein HSP27; spot protein MHC lp I Hỡnh 2.1 Phõn tỏch protein mụ gan ca bnh nhõn 8261 trờn bn gel 2-DE ii Hỡnh 2.2 Phõn tỏch protein mụ gan ca bnh nhõn 8935 trờn bn gel 2-DE Hỡnh 2.3 Phõn tỏch protein mụ gan ca bnh nhõn 8977 trờn bn gel 2-DE iii Hỡnh 2.4 Phõn tỏch protein mụ gan ca bnh nhõn 9242 trờn bn gel 2-DE Hỡnh 2.5 Phõn tỏch protein mụ gan ca bnh nhõn 8460 trờn bn gel 2-DE iv Hỡnh 2.6 Phõn tỏch protein mụ gan ca bnh nhõn 10480 trờn bn gel 2-DE v Ph lc Kt qu nhn dng protein bng c s d liu NCBI s dng phn mm Mascot vi vii Ph lc Kt qu phn ng Western Blot mu mụ gan ca bnh nhõn HCC A - Mụ gan bỡnh thng B - Mụ gan ung th Hỡnh 4.1 Kt qu phn ng Western Blot mu mụ gan ca bnh nhõn 9242 A - Mụ gan bỡnh thng B - Mụ gan ung th Hỡnh 4.2 Kt qu phn ng Western Blot mu mụ gan ca bnh nhõn 8935 A - Mụ gan bỡnh thng B - Mụ gan ung th Hỡnh 4.3 Kt qu phn ng Western Blot mu mụ gan ca bnh nhõn 8460 viii [...]... ung th gan nguyờn phỏt v ung th gan th phỏt (di cn) Ung th gan nguyờn phỏt l ung th xut phỏt t cỏc loi t bo gan (hỡnh 1) Ung th gan th phỏt l do cỏc t bo ung th ca cỏc c quan, b phn khỏc trong c th di chuyn theo dũng mỏu hoc mch bch huyt ti gan v nh khu ti õy, hỡnh thnh khi u Ung th di cn gan khụng mang cỏc c trng ca ung th xut phỏt t gan, vỡ vy, trong phm vi lun vn ny, chỳng tụi ch cp ti ung th gan. .. nht l ung th t bo biu mụ gan (Hepatocellular Carcinoma - HCC) thng gi l ung th gan [49] Hỡnh 1 Ung th gan xut phỏt t chớnh t bo gan [49] 1.1.1 Phõn loi ung th gan nguyờn phỏt Da vo phõn loi mụ bnh hc v loi t bo phỏt sinh ung th, ung th gan nguyờn phỏt c phõn thnh cỏc loi chớnh nh sau [42]: 3 Luận văn cao học - Lê Lan Ph-ơng Ung th t bo biu mụ gan l dng ung th biu mụ hay gp nht trong ung th gan Ung th... QUAN V UNG TH GAN Ung th gan l mt trong cỏc dng bnh lý ph bin nht trờn th gii v t l t vong do ung th gan xp hng th ba trờn tng s cỏc trng hp t vong do ung th Ti M, t nm 2004 n 2008, t l mc mi ung th gan mi nm tng 3,6% nam gii v 3% n gii, xu hng ny ó tn ti t nm 1992 Cỏc nh nghiờn cu d oỏn trong nm 2012 s cú khong 28.720 trng hp mc mi ung th gan, hn 80% trong s ú l ung th t bo biu mụ gan [8] Ung th gan. .. dch ca bnh nhõn ung th gan nhm mc ớch: - Tỡm hiu s biu hin ca cỏc protein mụ gan ung th so vi mụ gan bỡnh thng ca bnh nhõn ung th gan thụng qua cỏc im protein trờn bn gel in di hai chiu - Nhn dng c mt s protein khỏc bit c trng gia mụ gan ung th so vi mụ gan bỡnh thng - Xỏc nh c protein cú phn ng min dch (khỏng nguyờn ung th gan) c hiu vi cỏc khỏng th trong huyt tng bnh nhõn ung th gan ti c thc hin... ra l xỏc nh c 5000 protein gan, 5500 khung c m ca cỏc gene biu hin gan ngi c lu gi trong ngõn hng khung c m Bờn cnh ú, mt h thng hn 1000 tng tỏc gia protein - protein gan ngi c tỡm thy v miờu t Nhng thay i v proteomics trong mụ gan v huyt tng ó c nghiờn cu v so sỏnh trong quỏ trỡnh tin trin ca cỏc bnh v gan, t mụ gan bỡnh thng ti viờm gan, x gan, ung th gan v ung th gan di cn T nhng nghiờn cu ú,... õy l loi t bo gan ch yu, do ú, HCC chim 85 - 95% cỏc trng hp ung th gan nguyờn phỏt [32] - Ung th t bo ng mt, õy l dng ung th bt u cỏc ng mt nh trong gan Dng ung th ny ch chim khong 5 - 15% [32] Nguy c mc dng ung th ny s tng cao khi bnh nhõn b si mt, loột ng tiờu húa hay nhim cỏc dng ký sinh trựng trong gan - Ung th biu mụ hn hp, õy l dng ung th him gp, t bo ung th gm c t bo biu mụ gan v t bo ng mt... dựng sng lc ngi cú nguy c b ung th gan cao [39] Tuy vy, AFP trong mỏu khụng c trng cho ung th gan vỡ nng AFP cú th cng tng trong mt s trng hp khỏc: thai nghộn bỡnh thng, viờm gan giai on phc hi, viờm gan mn tớnh, ung th t bo mm nh ung th tinh hon hay mc mt s dng ung th khỏc nh ung th d dy, ung th ng mt [5] Xột nghim AFP cng khụng nhy bi cú khong 15% - 30% bnh nhõn ung th gan m hm lng AFP vn khụng tng... trng hp mc ung th gan v 695.900 ca t vong do ung th gan Vit Nam thuc khu vc ụng Nam l ni cú t l mc ung th gan cao nht [18] Theo thng kờ ca bnh vin K, nc ta cú t l mc ung th gan nguyờn phỏt cao do nhim virus viờm gan B v virus viờm gan C [41] Do ú, vic nghiờn cu v tỏc nhõn gõy bnh, c ch v phng phỏp chn oỏn, phỏt hin sm ung th gan cú vai trũ quan trng i vi vic gim t l mc v gim t l t vong loi ung th ny... cỏc protein biu hin khỏc bit ca mụ gan ung th v mụ gan lnh trờn cựng lỏ gan ca bnh nhõn ung th gan [6], ng thi cng tin hnh phõn tớch biu hin khỏc bit ca protein huyt tng ca bnh nhõn ung th gan s dng k thut in di hai chiu [36] Tip tc thc hin hng nghiờn cu proteomics ung th gan ti Vit Nam, trong ti ny, chỳng tụi ó kt hp phõn tớch proteomics mụ gan ca bnh nhõn ung th gan v thc hin phn ng min dch Western... bo ng mt - U nguyờn bo gan l loi u gan ỏc tớnh tr em, him gp ngi ln U nguyờn bo gan xut hin cú th do cỏc gene hot ng bt thng - Ung th t bo mch l loi ung th rt him gp, nú bt ngun t mch mỏu trong gan do gan tip xỳc vi cỏc húa cht cụng nghip nh vinyl chlorid hoc cỏc húa cht gõy ung th 1.1.2 Nguyờn nhõn dn ti ung th gan Mc dự vn cũn nhiu tranh cói v c ch chớnh xỏc dn n ung th gan, song cỏc nghiờn cu