• 1.四川大學(xué)華西醫(yī)院普外科(成都610041);;
  • 2.四川省雅安市人民醫(yī)院肝膽外科(雅安625000);

目的  構(gòu)建胰腺癌細(xì)胞生存蛋白(survivin)表達(dá)的RNA干擾(RNAi)抑制載體，并研究其對survivin表達(dá)的抑制作用。方法  用免疫熒光技術(shù)和RT-PCR檢測胰腺癌細(xì)胞PANC-1中survivin及其mRNA的表達(dá)，并把survivin基因克隆到T載體進(jìn)行測序； 構(gòu)建針對survivin缺失堿基基因和survivin基因的RNAi載體si-svv-1和si-svv-2，用脂質(zhì)體轉(zhuǎn)染胰腺癌細(xì)胞PANC-1后，用RT-PCR、DNA梯狀電泳及流式細(xì)胞術(shù)分析比較2種干擾載體對survivin mRNA表達(dá)的抑制情況和檢測細(xì)胞凋亡。結(jié)果  survivin在胰腺癌細(xì)胞PANC-1中高表達(dá)； si-svv-2對survivin mRNA的表達(dá)抑制率達(dá)(72.43±8.04)%，而si-svv-1為0； si-svv-2能誘導(dǎo)胰腺癌細(xì)胞PANC-1的凋亡, 72 h細(xì)胞凋亡率達(dá)（12.36±1.44）%。結(jié)論  本研究成功建立胰腺癌細(xì)胞survivin表達(dá)RNAi抑制載體，該載體可特異有效地抑制胰腺癌細(xì)胞PANC-1 survivin的表達(dá)并誘導(dǎo)胰腺癌細(xì)胞PANC-1凋亡。

引用本文： 何滿西,李耿,姚有貴,蒲道深,田伯樂,胡偉明,張肇達(dá). 胰腺癌細(xì)胞survivin表達(dá)RNAi抑制載體的構(gòu)建及其抑制作用的研究. 中國普外基礎(chǔ)與臨床雜志, 2008, 15(11): 829-834. doi: 復(fù)制

版權(quán)信息： ?四川大學(xué)華西醫(yī)院華西期刊社《中國普外基礎(chǔ)與臨床雜志》版權(quán)所有，未經(jīng)授權(quán)不得轉(zhuǎn)載、改編

1. Rubin P， Williams JP 主編． 孫 燕, 湯釗猷主譯. 臨床腫瘤學(xué) [M]. 第8版． 北京： 人民衛(wèi)生出版社， 2006∶768.
2. 陸星華． 胰腺癌研究現(xiàn)狀與展望 [J]. 美中醫(yī)學(xué)， 2005； 2（3）∶73.
3. MacKenzie MJ. Molecular therapy in pancreatic adenocarcinoma [J]. Lancet Oncol, 2004; 5(9)∶541.
4. Lieberman J, Song E, Lee SK, et al. Interfering with disease: opportunities and roadblocks to harnessing RNA interference [J]. Trends Mol Med, 2003; 9(9)∶397.
5. Olie RA, Simes-Wüst AP, Baumann B, et al. A novel antisense oligonucleotide targeting survivin expression induces apoptosis and sensitizes lung cancer cells to chemotherapy [J]. Cancer Res, 2000; 60(11)∶2805.
6. 何滿西， 孟令祥， 唐 勇， 等． RNAi抑制胰腺癌細(xì)胞survivin表達(dá)并誘導(dǎo)細(xì)胞凋亡的實(shí)驗(yàn)研究 [J]. 四川大學(xué)學(xué)報(bào)（醫(yī)學(xué)版）， 2006； 37（4）∶520.
7. Tamm I, Wang Y, Sausville E, et al. IAP-family protein survivin inhibits caspase activity and apoptosis induced by Fas (CD95), Bax, caspases, and anticancer drugs [J]. Cancer Res, 1998; 58(23)∶5315.
8. Islam A, Kageyama H, Takada N, et al. High expression of Survivin, mapped to 17q25, is significantly associated with poor prognostic factors and promotes cell survival in human neuroblastoma [J]. Oncogene, 2000; 19(5)∶617.
9. 李哲夫， 陳孝平， 張萬廣. 肝細(xì)胞癌合并門靜脈癌栓組織中PDGF和survivin表達(dá)的意義 [J]. 中國普外基礎(chǔ)與臨床雜志, 2007； 14（1）∶46.
10. 謝貽祥， 孟 剛. survivin在乳腺癌中的表達(dá)及其意義 [J]. 中國普外基礎(chǔ)與臨床雜志, 2007； 14（3）∶263.
11. Wang Z, Xie Y, Wang H. Changes in survivin messenger RNA level during chemotherapy treatment in ovarian cancer cells[J].Cancer Biol Ther, 2005; 4(7)∶716.
12. Van Valen F, Fulda S, Schfer KL, et al. Selective and nonselective toxicity of TRAIL/Apo2L combined with chemotherapy in human bone tumour cells vs. normal human cells [J]. Int J Cancer, 2003; 107(6)∶929.
13. 陳曦海， 張豈凡， 馬 榮. 反義survivin誘導(dǎo)胃癌SGC7901細(xì)胞凋亡及逆轉(zhuǎn)耐藥的研究 [J].中國普外基礎(chǔ)與臨床雜志, 2007； 14（1）∶63.
14. Satoh K, Kaneko K, Hirota M, et al. Expression of survivin is correlated with cancer cell apoptosis and is involved in the development of human pancreatic duct cell tumors [J]. Cancer, 2001; 92(2)∶271.
15. Jinfeng M, Kimura W, Sakurai F,et al. Histopathological study of intraductal papillary mucinous tumor of the pancreas: special reference to the roles of Survivin and p53 in tumorigenesis of IPMT [J]. Int J Gastrointest Cancer, 2002; 32(2-3)∶73.
16. 王亞利， 宋天保， 王西京， 等． Survivin mRNA反義寡核苷酸誘導(dǎo)胰腺癌細(xì)胞凋亡 [J]. 世界華人消化雜志， 2004； 12(8)∶1872.
17. 戴德堅(jiān), 陸才德, 賴日勇, 等． 靶向抑制survivin對胰腺癌細(xì)胞增殖和凋亡效應(yīng)的影響 [J]. 中華醫(yī)學(xué)雜志, 2004； 84（22）∶1894.
18. Lu B, Mu Y, Cao C, et al. Survivin as a therapeutic target for radiation sensitization in lung cancer [J]. Cancer Res, 2004; 64(8)∶2840.
19. Fulda S, Debatin KM. Sensitization for tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis by the chemopreventive agent resveratrol [J]. Cancer Res, 2004; 64(1)∶337.
20. 唐朝暉, 鄒聲泉, 余 彥, 等． 不同劑量的X射線輻照對胰腺癌細(xì)胞Survivin表達(dá)的影響 [J]. 臨床外科雜志， 2007； 15（5）∶328.
21. Williams NS, Gaynor RB, Scoggin S, et al. Identification and validation of genes involved in the pathogenesis of colorectal cancer using cDNA microarrays and RNA interference [J]. Clin Cancer Res, 2003; 9(3)∶931.
22. Fire A, Xu S, Montgomery MK, et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans[J]. Nature, 1998; 391(6669)∶806.
23. Miyagishi M, Taira K. Development and application of siRNA expression vector [J]. Nucleic Acids Res Suppl, 2002; 2(2)∶113.
  1. 1. Rubin P， Williams JP 主編． 孫 燕, 湯釗猷主譯. 臨床腫瘤學(xué) [M]. 第8版． 北京： 人民衛(wèi)生出版社， 2006∶768.
  2. 2. 陸星華． 胰腺癌研究現(xiàn)狀與展望 [J]. 美中醫(yī)學(xué)， 2005； 2（3）∶73.
  3. 3. MacKenzie MJ. Molecular therapy in pancreatic adenocarcinoma [J]. Lancet Oncol, 2004; 5(9)∶541.
  4. 4. Lieberman J, Song E, Lee SK, et al. Interfering with disease: opportunities and roadblocks to harnessing RNA interference [J]. Trends Mol Med, 2003; 9(9)∶397.
  5. 5. Olie RA, Simes-Wüst AP, Baumann B, et al. A novel antisense oligonucleotide targeting survivin expression induces apoptosis and sensitizes lung cancer cells to chemotherapy [J]. Cancer Res, 2000; 60(11)∶2805.
  6. 6. 何滿西， 孟令祥， 唐 勇， 等． RNAi抑制胰腺癌細(xì)胞survivin表達(dá)并誘導(dǎo)細(xì)胞凋亡的實(shí)驗(yàn)研究 [J]. 四川大學(xué)學(xué)報(bào)（醫(yī)學(xué)版）， 2006； 37（4）∶520.
  7. 7. Tamm I, Wang Y, Sausville E, et al. IAP-family protein survivin inhibits caspase activity and apoptosis induced by Fas (CD95), Bax, caspases, and anticancer drugs [J]. Cancer Res, 1998; 58(23)∶5315.
  8. 8. Islam A, Kageyama H, Takada N, et al. High expression of Survivin, mapped to 17q25, is significantly associated with poor prognostic factors and promotes cell survival in human neuroblastoma [J]. Oncogene, 2000; 19(5)∶617.
  9. 9. 李哲夫， 陳孝平， 張萬廣. 肝細(xì)胞癌合并門靜脈癌栓組織中PDGF和survivin表達(dá)的意義 [J]. 中國普外基礎(chǔ)與臨床雜志, 2007； 14（1）∶46.
  10. 10. 謝貽祥， 孟 剛. survivin在乳腺癌中的表達(dá)及其意義 [J]. 中國普外基礎(chǔ)與臨床雜志, 2007； 14（3）∶263.
  11. 11. Wang Z, Xie Y, Wang H. Changes in survivin messenger RNA level during chemotherapy treatment in ovarian cancer cells[J].Cancer Biol Ther, 2005; 4(7)∶716.
  12. 12. Van Valen F, Fulda S, Schfer KL, et al. Selective and nonselective toxicity of TRAIL/Apo2L combined with chemotherapy in human bone tumour cells vs. normal human cells [J]. Int J Cancer, 2003; 107(6)∶929.
  13. 13. 陳曦海， 張豈凡， 馬 榮. 反義survivin誘導(dǎo)胃癌SGC7901細(xì)胞凋亡及逆轉(zhuǎn)耐藥的研究 [J].中國普外基礎(chǔ)與臨床雜志, 2007； 14（1）∶63.
  14. 14. Satoh K, Kaneko K, Hirota M, et al. Expression of survivin is correlated with cancer cell apoptosis and is involved in the development of human pancreatic duct cell tumors [J]. Cancer, 2001; 92(2)∶271.
  15. 15. Jinfeng M, Kimura W, Sakurai F,et al. Histopathological study of intraductal papillary mucinous tumor of the pancreas: special reference to the roles of Survivin and p53 in tumorigenesis of IPMT [J]. Int J Gastrointest Cancer, 2002; 32(2-3)∶73.
  16. 16. 王亞利， 宋天保， 王西京， 等． Survivin mRNA反義寡核苷酸誘導(dǎo)胰腺癌細(xì)胞凋亡 [J]. 世界華人消化雜志， 2004； 12(8)∶1872.
  17. 17. 戴德堅(jiān), 陸才德, 賴日勇, 等． 靶向抑制survivin對胰腺癌細(xì)胞增殖和凋亡效應(yīng)的影響 [J]. 中華醫(yī)學(xué)雜志, 2004； 84（22）∶1894.
  18. 18. Lu B, Mu Y, Cao C, et al. Survivin as a therapeutic target for radiation sensitization in lung cancer [J]. Cancer Res, 2004; 64(8)∶2840.
  19. 19. Fulda S, Debatin KM. Sensitization for tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis by the chemopreventive agent resveratrol [J]. Cancer Res, 2004; 64(1)∶337.
  20. 20. 唐朝暉, 鄒聲泉, 余 彥, 等． 不同劑量的X射線輻照對胰腺癌細(xì)胞Survivin表達(dá)的影響 [J]. 臨床外科雜志， 2007； 15（5）∶328.
  21. 21. Williams NS, Gaynor RB, Scoggin S, et al. Identification and validation of genes involved in the pathogenesis of colorectal cancer using cDNA microarrays and RNA interference [J]. Clin Cancer Res, 2003; 9(3)∶931.
  22. 22. Fire A, Xu S, Montgomery MK, et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans[J]. Nature, 1998; 391(6669)∶806.
  23. 23. Miyagishi M, Taira K. Development and application of siRNA expression vector [J]. Nucleic Acids Res Suppl, 2002; 2(2)∶113.