33 6 2011 6 Journal of Ningxia Medical University 511 1674-6309 2011 06-0511 - 04 AnnexinA2 P19 1 2 3 3 3 1. 415700 2. 750004 3. 750004 AnnexinA2 P19 62 IHC RT - PCR Western blot AnnexinA2P19 mrna AnnexinA2 mrna P < 0. 05 P19 mrna P < 0. 05 AnnexinA2 P19 P > 0. 05 P < 0. 05 AnnexinA2 P19 mrna AnnexinA2 P19 R735. 7 A primary hepatic carcinoma PHC 5 1-3 AnnexinA2 1. 2 Annexin 62 45 17 34 ~ 81 62 HBsAg 47 4-6 > 5cm 36 5cm 26 Edmondson - steiner I 12 Ⅱ27 Ⅲ P19 G 1 CDK4 /6 G 1 /S 15 Ⅳ8 16 1. 3 RT - PCR AnnexinA2 p19 mrna 7-8 P19 IHC RT - PCR Western blot Trizol AnnexinA2 P19 mrna RNA RNA cdna Genebank 1 PCR 1. 1 mrna5μg 2009 5 - OligodNTP1. 0μL 20μL 70 5min 2010 8 5 5μL 10mmol L - 1 5cm 62 dntpmixtrue 1. 25μL 0. 5μL AMV 5min 0. 5cm 0. 5cm 0. 5cm 1. 0μL 2. 25μL 42 1h 70 10min PCR 50μL 10 Buffer 5. 0μL 2 5mL 10mmol L - 1 dntp 1. 0μL20mmol L - 1 2010-12 - 26 81060044 1979 - E - mail huangfeihong201314@yahoo. com. cn EP - 80 1. 0μL Taq 0. 5μL CDNA 1. 0μL 38. 5μL AnnexinA2 5 - ATT- GCCTTCGCCTACC - 3 5 - GCTCTTCTAC- CCTTTGC - 3 94 1min 56 1min 72 1min 30 329bp p19
512 33 5 - GGGGCACTTCCAAT - 3 5 - CAC- P < 0. 05 2 CTATAAGCCACAAAC - 3 94 30s 54 30s 72 2. 2 western 30s 319bp β - actin marker 62 5 - GCGCCTGGTCAC- AnnexinA2 P19 CAGGGCTGCTT - 3 5 - TACCGAAGTTGT- CATGGATGACCT - 3 533bp β - actin 0. 84 ± PCR 2% 0. 11 0. 32 ± 0. 08 AnnexinA2 mrna P < 0. 05 3 P19 1. 4 Western AnnexinA2 P19 β - actin 0. 28 ± 0. 13 0. 78 ± 0. 21 p19 P < 0. 05 4 BCA AnnexinA2 P19 1 300santa cruz 4 12h PBST 15min 3 1 3000 santa cruz 4 3h PBST 10min 3 ECL 5min bandscan 1 β - actin 1. 5 IHC AnnexinA2 P19 S - P AnnexinA2 P19 AEC 40 10 5 = / 100% 5 1 2 3 4 a b c d m marker 10% 2 RT - PCR 10% 1. 6 SPSS 11. 5 α = 0. 05 AnnexinA2 P19 mrna 3 Western blot t χ 2 AnnexinA2 P19 AnnexinA2 χ 2 2 AnnexinA2 1 2 a b m marker RT - PCR AnnexinA2 mrna P19 mrna a b 2. 1 RT - PCR a b maker 62 4 Western blot AnnexinA2 P19 mrna P19 AnnexinA2 mrna 2. 3 IHC β - actin AnnexinA2 0. 90 ± 0. 14 0. 19 ± 0. 05 5 2 AnnexinA2 mrna P < 71. 0% 0. 05 1 P19 mrna 44 /62 14. 5% 9 /62 χ 2 = β - actin 0. 32 ± 0. 07 40. 367P < 0. 05 P19 0. 87 ± 0. 11 p19 mrna 6 2
6. AnnexinA2 P19 513 25. 8% 16 /62 66. 1% 41 / 62 χ 2 = 20. 293P < 0. 05 AnnexinA2 P19 AnnexinA2 P19 2. 4 AnnexinA2 P19 P > 0. 05 P < 62 0. 05 1 1 50 > 50 HBsAg > 5cm 5cm Ⅰ Ⅱ Ⅲ Ⅳ 3 45 17 27 35 47 15 36 26 39 23 16 46 AnnexinA2 P19 AnnexinA2 % χ2 P 33 73. 3 11 64. 7 19 70. 4 25 71. 4 38 80. 9 6 40. 0 26 72. 2 18 29. 2 24 61. 5 20 87. 0 15 93. 8 29 63. 0 0. 125 0. 723 0. 008 0. 927 7. 334 0. 007 0. 066 0. 798 4. 537 0. 033 4. 044 0. 044 P19 % χ2 P 14 31. 1 2 11. 8 10 37. 0 6 17. 1 6 12. 8 10 66. 7 9 25. 0 7 26. 9 15 38. 5 1 4. 4 0 0. 0 16 34. 8 1. 507 0. 220 3. 151 0. 076 14. 554 0. 000 0. 029 0. 864 8. 794 0. 003 5. 794 0. 016 HeLa DNA AnnexinA2 mrna G 1 PHC S 4-5 AnnexinA2 G 1 - S S - G 2 9 AnnexinA2 5 AnnexinA2 AnnexinA2 An- nexina2 P19 AnnexinA2 P53 P53 P53 MDM2 10 - HBsAg AnnexinA2 12 HBsAg P19 AnnexinA2 P19 AnnexinA2 mrnp c - myc AnnexinA2 P19 DNA Paliwal Kalinichenko P19 α AnnexinA2 P19
514 33 G 1 /S binds to annexin2 in pancreatic cancer cells and pro- pancreatic cancer cell motility J. British Jour- motes 7-8 13 P19 nal of Cancer 2009 101 1145-1154. 7Chen Z Carracedo A Lin HK et al. Differential p53 - independent outcomes of p19arf loss in oncogenesis J. Sci Signal 2009 2 84 44. 8Ulanet DB Hanahan D. Loss of p19 Arf facilitates the angiogenic switch and tumor initiation in a multi - stage AnnexinA2 P19 cancer model via p53 - dependent and independent mechanisms J. Plos One 2010 5 8 1-10. 9. A2 J. 200830 1Hoshida Y Villanueva A Kobayashi M et al. Gene expression in fixed tissues and outcome in hepatocellular carcinoma J. N Engl J Med2008359 19 1995-2004. 2Llovet JM Bruix J. Molecular targeted therapies in hepatocellular carcinomaj. Hepatology 2008 48 4 1312-1327. 3Zulehner G Mikula M Schneller D et al. Nuclear beta - catenin induces an early liver progenitor phenotype in hepatocellular carcinoma and promotes tumor recurrence J. Am J Pathol 2010 176 1 472-481. 4Huang Y Jin Y Yan CH et al. Involvement of AnnexinA2 in P53 induced apoptosis in lung cancerj. Mol Cell Biochem 2008 309 117-123. 13Sebastian T Johnson PF. RasV12 - mediated down - regulation of CCAAT / enhancer binding protein beta in immortalized fibroblasts requires loss of p19arf and fa- 5. Annexin A2 cilitates bypass of oncogene - induced senescence J. J. 2007 36 2 129-132. Cancer Res 2009 69 6 2588-2598. 6Nedjadi T Kitteringham N Campbell F et al. S100A6 307-311. 10Chen YW Paliwal S Draheim K et al. p19arf inhibits the invasion of hepatocellular carcinoma cells by binding to C - terminal binding proteinj. Cancer Res 2008 68 2 476-482. 11Liu L Li J Xu YP et a1. RNA silencing suppressor p19 Regulates the expressions of cell cycle related genesj. Progress in Biochemistry and Biophysics 2009 36 5 541-548. 12Camats M Kokolo M Heesom KJ et al. P19 H - ras induces G1/S phase delay maintaining cells in a reversible quiescence state J. Plos One 2009 4 12 1-10. Expression and Significance of AnnexinA2 and P19 in Patients with Primary Hepatic Carcinoma HUANG Bo 1 2 LU Zhen - hui 3 SONG Jian - jun 3 ZHAO Guo - zhong 3 1. Taoyuan First People s Hospital Taoyuan 415700 2. Ningxia Medical UniversityYinchuan 750004 3. Dept. of Hepatobiliary Surgery the General Hospital of Ningxia Med. Univ. Yinchuan 750004 Abstract Objective To detect the expression of AnnexinA2 and P19 in patients with primary hepatic carcinoma and to explore the effect of their expression in occurrence development and metastasis of primary hepatic carcinoma. Methods RT - PCR Western blot and IHC techniques were used to detect the expression of AnnexinA2 and P19 in patients with primary hepatic carcinoma. Results AnnexinA2 and P19 expression in primary hepatic carcinoma tissues were significantly different from those in the adjacent liver tissues. The expression of AnnexinA2 in the primary hepatic carcinoma tissues was significantly higher than that in the adjacent liver tissues P < 0. 05. The expression of P19 in the primary hepatic carcinoma tissue was significantly lower than that in the adjacent liver tissues P < 0. 05. Conclusion AnnexinA2 and P19 expression has important value for prediction of occurrence and metastasis. They maybe supply evidence for clinical therapy and judgement of prognosis. Key words primary hepatic carcinoma AnnexinA2 P19