• 廣州醫(yī)學(xué)院第一附屬醫(yī)院肝膽外科（廣東廣州 510120）;

目的觀察靶向性重組腺病毒Ad-EAFP-PALB/r-Caspase-3對(duì)原發(fā)性肝癌HepG2細(xì)胞及裸鼠皮下種植瘤的治療作用。方法采用先前構(gòu)建的靶向性重組腺病毒Ad-EAFP-PALB/r-Caspase-3感染肝癌HepG 2細(xì)胞、乳腺癌MDA-MB-231細(xì)胞以及正常肝L-02細(xì)胞，觀察細(xì)胞形態(tài)學(xué)改變，并通過流式細(xì)胞術(shù)檢測(cè)細(xì)胞凋亡指數(shù); 采用裸鼠皮下種植瘤模型、重組腺病毒瘤內(nèi)給藥治療，觀察腫瘤生長(zhǎng)情況及病理學(xué)改變。 結(jié)果流式細(xì)胞學(xué)可見HepG 2細(xì)胞凋亡和明顯的凋亡峰，MDA-MB-231和L-02細(xì)胞凋亡指數(shù)分別為0和7.3%，明顯低于HepG2細(xì)胞（48.2%），P lt;0.01。 種植腫瘤細(xì)胞2周后HepG 2細(xì)胞組、MDA-MB-231細(xì)胞組和對(duì)照組腫瘤體積大小分別為(0.26±0.31) cm 3、(0.25±0.15) cm 3和(0.26±0.28) cm 3，各組間差異無統(tǒng)計(jì)學(xué)意義（P gt;0.05）; 種植腫瘤細(xì)胞4周后HepG 2細(xì)胞組、MDA-MB-231細(xì)胞組和對(duì)照組腫瘤體積大小分別為(0.53±0.12) cm 3、(0.49±0.22) cm 3和(0.54±0.13) cm 3，各組間差異無統(tǒng)計(jì)學(xué)意義（P gt;0.05）; HepG 2細(xì)胞組腫瘤經(jīng)重組腺病毒Ad-EAFP-PALB/r-Caspase-3治療后8周時(shí)腫瘤體積為(0.65±0.13) cm 3，明顯小于MDA-MB-231細(xì)胞組〔(1.27±0.32) cm 3〕和對(duì)照組〔(1.43±1.09) cm 3〕，P lt;0.01，而后兩組之間比較差異無統(tǒng)計(jì)學(xué)意義（P gt;0.05）。 治療后HepG 2細(xì)胞組腫瘤細(xì)胞出現(xiàn)壞死區(qū)，伴淋巴浸潤(rùn)細(xì)胞和凋亡細(xì)胞; 對(duì)照組腫瘤細(xì)胞區(qū)域內(nèi)未見明顯壞死區(qū); 而MDA-MB-231細(xì)胞組腫瘤細(xì)胞呈實(shí)性排列，呈小腺腔樣結(jié)構(gòu)，異型性明顯，瘤巨細(xì)胞及多核瘤細(xì)胞可見，治療前、后未見明顯變化。 結(jié)論 靶向性重組腺病毒Ad-EAFP-PALB/r-Caspase-3具有有效靶向性凋亡誘導(dǎo)作用，體內(nèi)、外治療原發(fā)性肝癌有效，可能為原發(fā)性肝癌的靶向性基因治療提供新途徑。

引用本文： 王煒,沈浩元,劉猛. 靶向性重組腺病毒對(duì)裸鼠肝癌種植瘤的治療作用. 中國(guó)普外基礎(chǔ)與臨床雜志, 2011, 18(7): 722-727. doi: 復(fù)制

1. 王煒, 程衛(wèi), 丘昶儒. 肝細(xì)胞肝癌靶向性rCaspase3重組腺病毒的構(gòu)建與表達(dá) [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2008, 15(8): 589594.
2. 王煒, 沈浩元, 丘昶儒, 等. 靶向性反向半胱氨酸天冬氨酸蛋白酶3重組腺病毒誘導(dǎo)肝癌細(xì)胞凋亡的實(shí)驗(yàn)研究 [J]. 中華生物醫(yī)學(xué)工程雜志, 2010, 16(4): 298303.
3. 陳汝福, 孫宏武, 鄒倩. 反義硫代寡脫氧核苷酸抑制裸鼠人膽管癌生長(zhǎng)的研究 [J]. 中華實(shí)驗(yàn)外科雜志, 2000, 17(1): 2223.
4. 畢允力, 高解春, 許德華. 5氟胞嘧啶對(duì)表達(dá)胞嘧啶脫氨酶的人腎母細(xì)胞瘤裸鼠異種移植瘤的療效 [J]. 中華腫瘤雜志, 2000, 22(2): 113115.
5. 陳孝平, 張萬廣. 原發(fā)性肝癌面臨的難點(diǎn)與對(duì)策 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2008, 25(4): 231233.
6. Araida T, Higuchi R, Hamano M, et al. Hepatic resection in 485 R0 pT2 and pT3 cases of advanced carcinoma of the gallbladder: results of a Japanese Society of Biliary Surgery survey—a multicenter study [J]. Hepatobiliary Pancreat Surg, 2009, 16(2): 204215.
7. Cescon M, Cucchetti A, Grazi GL, et al. Indication of the extent of hepatectomy for hepatocellular carcinoma on cirrhosis by a simple algorithm based on preoperative variables [J]. Arch Surg, 2009, 144(1): 5763.
8. Ishizawa T, Hasegawa K, Kokudo N, et al. Risk factors and management of ascites after liver resection to treat hepatocellular carcinoma [J]. Arch Surg, 2009, 144(1): 4651.
9. Cho JY, Han HS, Yoon YS, et al. Outcomes of laparoscopic liver resection for lesions located in the right side of the liver [J]. Arch Surg, 2009, 144(1): 2529.
10. Miyazawa M, Aikawa M, Okada K, et al. Extended left hepatic lobectomy for hepatic hilar bile duct cancer: a novel surgical procedure in which the right hepatic duct is transected before the hepatoduodenal ligament is skeletonized [J]. Dig Surg, 2009, 26(1): 1519.
11. Smerdou C, Menne S, HernandezAlcoceba R, et al. Gene therapy for HCV/HBVinduced hepatocellular carcinoma [J]. Curr Opin Investig Drugs, 2010, 11(12): 13681377.
12. Sharapova OA, Pozdnykova NV, Laurinavichyute DK, et al. Highefficient expression, refolding and purification of functional recombinant Cterminal fragment of human alphafetoprotein [J]. Protein Expr Purif, 2010, 73(1): 3135.
13. Lam PY, Sia KC, Khong JH, et al. An efficient and safe herpes simplex virus type 1 amplicon vector for transcriptionally targeted therapy of human hepatocellular carcinomas [J]. Mol Ther, 2007, 15(6): 11291136.
14. Nussbaum T, Samarin J, Ehemann V, et al. Autocrine insulinlike growth factorII stimulation of tumor cell migration is a progression step in human hepatocarcinogenesis [J]. Hepatology, 2008, 48(1): 146156.
15. Sadeghi H, Hitt MM. Transcriptionally targeted adenovirus vectors [J]. Curr Gene Ther, 2005, 5(4): 411427.
16. Kanai F. Transcriptional targeted gene therapy for hepatocellular carcinoma by adenovirus vector [J]. Mol Biotechnol, 2001, 18(3): 243250.
17. Srinivasula SM, Ahmad M, MacFarlane M, et al. Generation of constitutively active recombinant caspases3 and 6 by rearrangement of their subunits [J]. J Biol Chem, 1998, 273(17): 1010710111.
18. 王煒, 秦兆寅, 王劍利, 等. 重組的rCaspase3促凋亡基因體內(nèi)外治療胰腺癌的試驗(yàn)研究 [J]. 中華肝膽外科雜志, 2003, 9(5): 303305.
19. 王煒, 秦兆寅, 劉志國(guó). 重組型Caspase3基因的構(gòu)建及其在胰腺癌細(xì)胞中凋亡活性的觀察 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2002, 9(4): 249251.
20. Park HJ, Kim MJ, Ha E, et al. Apoptotic effect of hesperidin through caspase3 activation in human colon cancer cells, SNUC4 [J]. Phytomedicine, 2008, 15(12): 147151.
21. Mouratidis PX, Colston KW, Dalgleish AG. Doxycycline induces caspasedependent apoptosis in human pancreatic cancer cells [J]. Int J Cancer, 2007, 120(4): 743752.
22. Végran F, Boidot R, Oudin C, et al. Overexpression of caspase3s splice variant in locally advanced breast carcinoma is associated with poor response to neoadjuvant chemotherapy [J]. Clin Cancer Res, 2006, 12(19): 57945800.
23. Aquino Esperanza JA, Aguirre MV, Aispuru GR, et al. In vivo 5fluorouracil[corrected] induced apoptosis on murine thymocytes: involvement of FAS, Bax and Caspase3 [J]. Cell Biol Toxicol, 2008, 24(5): 411422.
24. 易石堅(jiān), 李蘭蘭, 鐘德許, 等. 裸鼠皮下人肝癌種植瘤模型的建立 [J]. 中國(guó)醫(yī)藥導(dǎo)報(bào), 2008, 21(5): 2728.
25. 李立, 吳沛宏, 黃嘉凌, 等. 重組人內(nèi)皮抑素腺病毒抑制肝癌裸鼠移植瘤生長(zhǎng) [J]. 中華肝臟病雜志, 2008, 16(9): 542545.
26. Kurio N, Shimo T, Fukazawa T, et al. Antitumor effect in human breast cancer by TAE226, a dual inhibitor for FAK and IGFIR in vitro and in vivo [J]. Exp Cell Res, 2011, 317(8): 11341146.
  1. 1. 王煒, 程衛(wèi), 丘昶儒. 肝細(xì)胞肝癌靶向性rCaspase3重組腺病毒的構(gòu)建與表達(dá) [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2008, 15(8): 589594.
  2. 2. 王煒, 沈浩元, 丘昶儒, 等. 靶向性反向半胱氨酸天冬氨酸蛋白酶3重組腺病毒誘導(dǎo)肝癌細(xì)胞凋亡的實(shí)驗(yàn)研究 [J]. 中華生物醫(yī)學(xué)工程雜志, 2010, 16(4): 298303.
  3. 3. 陳汝福, 孫宏武, 鄒倩. 反義硫代寡脫氧核苷酸抑制裸鼠人膽管癌生長(zhǎng)的研究 [J]. 中華實(shí)驗(yàn)外科雜志, 2000, 17(1): 2223.
  4. 4. 畢允力, 高解春, 許德華. 5氟胞嘧啶對(duì)表達(dá)胞嘧啶脫氨酶的人腎母細(xì)胞瘤裸鼠異種移植瘤的療效 [J]. 中華腫瘤雜志, 2000, 22(2): 113115.
  5. 5. 陳孝平, 張萬廣. 原發(fā)性肝癌面臨的難點(diǎn)與對(duì)策 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2008, 25(4): 231233.
  6. 6. Araida T, Higuchi R, Hamano M, et al. Hepatic resection in 485 R0 pT2 and pT3 cases of advanced carcinoma of the gallbladder: results of a Japanese Society of Biliary Surgery survey—a multicenter study [J]. Hepatobiliary Pancreat Surg, 2009, 16(2): 204215.
  7. 7. Cescon M, Cucchetti A, Grazi GL, et al. Indication of the extent of hepatectomy for hepatocellular carcinoma on cirrhosis by a simple algorithm based on preoperative variables [J]. Arch Surg, 2009, 144(1): 5763.
  8. 8. Ishizawa T, Hasegawa K, Kokudo N, et al. Risk factors and management of ascites after liver resection to treat hepatocellular carcinoma [J]. Arch Surg, 2009, 144(1): 4651.
  9. 9. Cho JY, Han HS, Yoon YS, et al. Outcomes of laparoscopic liver resection for lesions located in the right side of the liver [J]. Arch Surg, 2009, 144(1): 2529.
  10. 10. Miyazawa M, Aikawa M, Okada K, et al. Extended left hepatic lobectomy for hepatic hilar bile duct cancer: a novel surgical procedure in which the right hepatic duct is transected before the hepatoduodenal ligament is skeletonized [J]. Dig Surg, 2009, 26(1): 1519.
  11. 11. Smerdou C, Menne S, HernandezAlcoceba R, et al. Gene therapy for HCV/HBVinduced hepatocellular carcinoma [J]. Curr Opin Investig Drugs, 2010, 11(12): 13681377.
  12. 12. Sharapova OA, Pozdnykova NV, Laurinavichyute DK, et al. Highefficient expression, refolding and purification of functional recombinant Cterminal fragment of human alphafetoprotein [J]. Protein Expr Purif, 2010, 73(1): 3135.
  13. 13. Lam PY, Sia KC, Khong JH, et al. An efficient and safe herpes simplex virus type 1 amplicon vector for transcriptionally targeted therapy of human hepatocellular carcinomas [J]. Mol Ther, 2007, 15(6): 11291136.
  14. 14. Nussbaum T, Samarin J, Ehemann V, et al. Autocrine insulinlike growth factorII stimulation of tumor cell migration is a progression step in human hepatocarcinogenesis [J]. Hepatology, 2008, 48(1): 146156.
  15. 15. Sadeghi H, Hitt MM. Transcriptionally targeted adenovirus vectors [J]. Curr Gene Ther, 2005, 5(4): 411427.
  16. 16. Kanai F. Transcriptional targeted gene therapy for hepatocellular carcinoma by adenovirus vector [J]. Mol Biotechnol, 2001, 18(3): 243250.
  17. 17. Srinivasula SM, Ahmad M, MacFarlane M, et al. Generation of constitutively active recombinant caspases3 and 6 by rearrangement of their subunits [J]. J Biol Chem, 1998, 273(17): 1010710111.
  18. 18. 王煒, 秦兆寅, 王劍利, 等. 重組的rCaspase3促凋亡基因體內(nèi)外治療胰腺癌的試驗(yàn)研究 [J]. 中華肝膽外科雜志, 2003, 9(5): 303305.
  19. 19. 王煒, 秦兆寅, 劉志國(guó). 重組型Caspase3基因的構(gòu)建及其在胰腺癌細(xì)胞中凋亡活性的觀察 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2002, 9(4): 249251.
  20. 20. Park HJ, Kim MJ, Ha E, et al. Apoptotic effect of hesperidin through caspase3 activation in human colon cancer cells, SNUC4 [J]. Phytomedicine, 2008, 15(12): 147151.
  21. 21. Mouratidis PX, Colston KW, Dalgleish AG. Doxycycline induces caspasedependent apoptosis in human pancreatic cancer cells [J]. Int J Cancer, 2007, 120(4): 743752.
  22. 22. Végran F, Boidot R, Oudin C, et al. Overexpression of caspase3s splice variant in locally advanced breast carcinoma is associated with poor response to neoadjuvant chemotherapy [J]. Clin Cancer Res, 2006, 12(19): 57945800.
  23. 23. Aquino Esperanza JA, Aguirre MV, Aispuru GR, et al. In vivo 5fluorouracil[corrected] induced apoptosis on murine thymocytes: involvement of FAS, Bax and Caspase3 [J]. Cell Biol Toxicol, 2008, 24(5): 411422.
  24. 24. 易石堅(jiān), 李蘭蘭, 鐘德許, 等. 裸鼠皮下人肝癌種植瘤模型的建立 [J]. 中國(guó)醫(yī)藥導(dǎo)報(bào), 2008, 21(5): 2728.
  25. 25. 李立, 吳沛宏, 黃嘉凌, 等. 重組人內(nèi)皮抑素腺病毒抑制肝癌裸鼠移植瘤生長(zhǎng) [J]. 中華肝臟病雜志, 2008, 16(9): 542545.
  26. 26. Kurio N, Shimo T, Fukazawa T, et al. Antitumor effect in human breast cancer by TAE226, a dual inhibitor for FAK and IGFIR in vitro and in vivo [J]. Exp Cell Res, 2011, 317(8): 11341146.