凋亡抑制因子survivin mRNA在鼻咽癌组织中的定量表达
摘要
目的 探讨survivin mRNA 在鼻咽癌组织中的表达及其与临床病理特征的关系。
方法 采用实时荧光定量PCR(FQ-PCR)法，检测46例鼻咽癌组织及18例正常鼻咽组织中survivin mRNA的表达。根据FQ-PCR以Ct值为标准，所得数值与实际表达量对数值呈反比的原理，以survivin mRNA Ct/GAPDH Ct的比值来评价不同组织survivin mRNA的表达。
结果 survivin mRNA在鼻咽癌组织中的表达为1.037±0.133，在正常鼻咽组织中的表达为1.187±0.092；统计分析差异显著意义（t=4.03、P=0.000）。survivin mRNA表达与鼻咽癌TNM分期、淋巴结转移、性别和年龄的关系无明显统计学意义（P均>0.5）。
结论 survivin mRNA在鼻咽癌中的表达高于正常鼻咽组织，在鼻咽癌的发生发展中可能起重要作用。实时荧光定量PCR用于检测survivin，具有实时检测、重复性好、灵敏度高、特异性高的特点。
Objective To study the expression of survivin mRNA and the relationship between survivin mRNA and clinical pathological features in Nasopharyngeal carcinoma (NPC).
Methods The expression of survivin mRNA in 46 samples of nasopharyngeal carcinoma tissues and 18 samples of normal nasopharyngeal tissues were detected by FQ-PCR,using glyceraldehydes-3-phophate dehydrogenase (GAPDH) as internal standard. On the principle that the CT value is in inverse proportion to the log value of original copy number of target sequence, SURVIVIN CT/ GAPDH CT was used to evaluate the mRNA expression level in different tissues.
Results The survivin mRNA levels in nasopharyngeal carcinoma tissues (1.037±0.133) was significantly higher than those in normal nasopharyngeal tissues (1.187±0.092), (P=0.000).By statistical analysis , there was no relationship between survivin mRNA and sex, age, TNM stages, lymph node metastases(P>0.05).
Conclusions The expression of survivin mRNA is higher in nasopharyngeal carcinoma tissues than in normal nasopharyngeal tissues. The expression of survivin mRNA in nasopharyngeal carcinoma may play an important role
in tumorigenesis through inhibition of apoptosis. FQ-PCR has the characteristics of the real-time detection、good repetition、high sensitivity and high specificity through detecting survivin in NPC.
方法 采用实时荧光定量PCR(FQ-PCR)法，检测46例鼻咽癌组织及18例正常鼻咽组织中survivin mRNA的表达。根据FQ-PCR以Ct值为标准，所得数值与实际表达量对数值呈反比的原理，以survivin mRNA Ct/GAPDH Ct的比值来评价不同组织survivin mRNA的表达。
结果 survivin mRNA在鼻咽癌组织中的表达为1.037±0.133，在正常鼻咽组织中的表达为1.187±0.092；统计分析差异显著意义（t=4.03、P=0.000）。survivin mRNA表达与鼻咽癌TNM分期、淋巴结转移、性别和年龄的关系无明显统计学意义（P均>0.5）。
结论 survivin mRNA在鼻咽癌中的表达高于正常鼻咽组织，在鼻咽癌的发生发展中可能起重要作用。实时荧光定量PCR用于检测survivin，具有实时检测、重复性好、灵敏度高、特异性高的特点。
Objective To study the expression of survivin mRNA and the relationship between survivin mRNA and clinical pathological features in Nasopharyngeal carcinoma (NPC).
Methods The expression of survivin mRNA in 46 samples of nasopharyngeal carcinoma tissues and 18 samples of normal nasopharyngeal tissues were detected by FQ-PCR,using glyceraldehydes-3-phophate dehydrogenase (GAPDH) as internal standard. On the principle that the CT value is in inverse proportion to the log value of original copy number of target sequence, SURVIVIN CT/ GAPDH CT was used to evaluate the mRNA expression level in different tissues.
Results The survivin mRNA levels in nasopharyngeal carcinoma tissues (1.037±0.133) was significantly higher than those in normal nasopharyngeal tissues (1.187±0.092), (P=0.000).By statistical analysis , there was no relationship between survivin mRNA and sex, age, TNM stages, lymph node metastases(P>0.05).
Conclusions The expression of survivin mRNA is higher in nasopharyngeal carcinoma tissues than in normal nasopharyngeal tissues. The expression of survivin mRNA in nasopharyngeal carcinoma may play an important role
in tumorigenesis through inhibition of apoptosis. FQ-PCR has the characteristics of the real-time detection、good repetition、high sensitivity and high specificity through detecting survivin in NPC.
引文
Deveraux,Q.L,Takahashi,R,Salvesen,G.S,et al.Nature 1997, 388: 300-304.
LaCasse EC, Baird S, Korneluk RG,et al. The inhibitors of apoptosis (IAPs) and their emerging role in cancer. Oncogene, 1998 ,17(25):3247-3259.
3. Sela B.Survivin:anti-apoptosis protein and a prognostic marker for tumor progression and recurrence. Harefuah, 2002,141(1): 103-107.
4. 闵华庆,郭翔.鼻咽癌研究回顾与展望.国外医学：肿瘤学分册.2000,27(1)：11-14.
5. Ikeguchi M, Hirooka Y, Kaibara N. Quantitative analysis of apoptosis-related gene expression in hepatocellular carcinoma. Cancer. 2002 Nov 1;95(9):1938-1945.
6. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001 Dec;25(4):402-408.
7. 陈意生,史景泉主编.肿瘤分子细胞生物学.第1版.北京：人民军医出版社,2002.152-154.
8. Jiang X, Wilford C, Duensing S，et al. Participation of Survivin in mitotic and apoptotic activities of normal and tumor- derived cells. J Cell Biochem. 2001 Aug 1-9; 83(2):342-354.
9. Ambrosini G,Adida C,Altieri DC. A novel anti-apoptosis gene,
survivin, expressed in cancer and lymphoma. Nat Med, 1997,3(8):917-921.
10. Li F, Ambrosini G,Chu EY,et al. Control of apoptosis and mitotic spindle checkpoint by survivin. Nature,1998, 396(6711):580-4.
11. Andrade F,Roy S,Micholson D,et al.Granzyme directly and efficiently cleaves several downstream caspase substrates: implications for CTL-induced apoptosis [J].Immunity,1998, 8(4):451-460.
12. 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. Cancer Res.1998;58 (23): 5315-5320.
13. Shinozawa I,Inokuchi K,Wakabayashi I,et al.Disturbed expre- ssion of the anti-apoptosis gene, survivin, and EPR-1 in hematological malignancies. Leuk Res. 2000 Nov;24(11):965-70.
14. Bao R, Connolly DC, Murphy M，et al. Activation of cancer- specific gene expression by the survivin promoter. J Natl Cancer Inst. 2002 Apr 3;94(7):522-528.
15.韦宜峰,黄光武. 实时荧光定量PCR在鼻咽癌中的应用.中国医学文摘·肿瘤学,2003,17(2):168-170.
16. Wall SJ,Edward DR.Quantitative reverse transcription-polyme- rase chain reaction (TR-PCR):A comparison of prime-dropping ,
competitive,and real-time RT-PCRs.Anal Biochem,2002, 300(2): 269-273.
17. Bustin S.A. Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J. Mol. Endocrinol,2000,25:169-193.
18. Ikeguchi M, Kaibara N. survivin messenger RNA expression is a good prognostic biomarker for oesophageal carcinoma. Br J Cancer, 2002 ,87(8):883-887.
19. Miyachi K, Sasaki K, Onodera S, et al. Correlation between survivin mRNA expression and lymph node metastasis in gastric cancer. Gastric Cancer. 2003;6(4):217-224.
20. Kato J, Kuwabara Y, Mitani M, et al. Expression of survivin in esophageal cancer: correlation with the prognosis and response to chemotherapy. Int J Cancer. 2001 Mar 20;95(2): 92-95.
21. Kaur P, Kallakury BS, Sheehan CE, et al. Survivin and Bcl-2 expression in prostatic adenocarcinomas. Arch Pathol Lab Med. 2004 Jan;128(1):39-43 .
22. Ku JH, Kwak C, Lee HS, et al. Expression of survivin, a novel inhibitor of apoptosis, in superficial transitional cell carcinoma of the bladder. J Urol. 2004 Feb;171(2 Pt 1):631-635.
23. Grabowski P, Kuhnel T, Muhr-Wilkenshoff F, et al. Prognostic value of nuclear survivin expression in oesophageal squamous cell
carcinoma. Br J Cancer. 2003 Jan 13;88(1):115-119.
24. Sarela AI, Macadam RC, Farmery SM, et al . Expression of the antiapoptosis gene, survivin, predicts death from recurrent colorectal carcinoma. Gut. 2000 May;46(5):645-650.
25. Grossman D, McNiFF JM, Li F, et al. Expression of the apoptosis inhibitor, survivin，in nonmelanoma skin cancer and targeting in a keratincyte cell line . Lab Invest , 1999,79:1121-1126.
LaCasse EC, Baird S, Korneluk RG,et al. The inhibitors of apoptosis (IAPs) and their emerging role in cancer. Oncogene, 1998 ,17(25):3247-3259.
3. Sela B.Survivin:anti-apoptosis protein and a prognostic marker for tumor progression and recurrence. Harefuah, 2002,141(1): 103-107.
4. 闵华庆,郭翔.鼻咽癌研究回顾与展望.国外医学：肿瘤学分册.2000,27(1)：11-14.
5. Ikeguchi M, Hirooka Y, Kaibara N. Quantitative analysis of apoptosis-related gene expression in hepatocellular carcinoma. Cancer. 2002 Nov 1;95(9):1938-1945.
6. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001 Dec;25(4):402-408.
7. 陈意生,史景泉主编.肿瘤分子细胞生物学.第1版.北京：人民军医出版社,2002.152-154.
8. Jiang X, Wilford C, Duensing S，et al. Participation of Survivin in mitotic and apoptotic activities of normal and tumor- derived cells. J Cell Biochem. 2001 Aug 1-9; 83(2):342-354.
9. Ambrosini G,Adida C,Altieri DC. A novel anti-apoptosis gene,
survivin, expressed in cancer and lymphoma. Nat Med, 1997,3(8):917-921.
10. Li F, Ambrosini G,Chu EY,et al. Control of apoptosis and mitotic spindle checkpoint by survivin. Nature,1998, 396(6711):580-4.
11. Andrade F,Roy S,Micholson D,et al.Granzyme directly and efficiently cleaves several downstream caspase substrates: implications for CTL-induced apoptosis [J].Immunity,1998, 8(4):451-460.
12. 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. Cancer Res.1998;58 (23): 5315-5320.
13. Shinozawa I,Inokuchi K,Wakabayashi I,et al.Disturbed expre- ssion of the anti-apoptosis gene, survivin, and EPR-1 in hematological malignancies. Leuk Res. 2000 Nov;24(11):965-70.
14. Bao R, Connolly DC, Murphy M，et al. Activation of cancer- specific gene expression by the survivin promoter. J Natl Cancer Inst. 2002 Apr 3;94(7):522-528.
15.韦宜峰,黄光武. 实时荧光定量PCR在鼻咽癌中的应用.中国医学文摘·肿瘤学,2003,17(2):168-170.
16. Wall SJ,Edward DR.Quantitative reverse transcription-polyme- rase chain reaction (TR-PCR):A comparison of prime-dropping ,
competitive,and real-time RT-PCRs.Anal Biochem,2002, 300(2): 269-273.
17. Bustin S.A. Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J. Mol. Endocrinol,2000,25:169-193.
18. Ikeguchi M, Kaibara N. survivin messenger RNA expression is a good prognostic biomarker for oesophageal carcinoma. Br J Cancer, 2002 ,87(8):883-887.
19. Miyachi K, Sasaki K, Onodera S, et al. Correlation between survivin mRNA expression and lymph node metastasis in gastric cancer. Gastric Cancer. 2003;6(4):217-224.
20. Kato J, Kuwabara Y, Mitani M, et al. Expression of survivin in esophageal cancer: correlation with the prognosis and response to chemotherapy. Int J Cancer. 2001 Mar 20;95(2): 92-95.
21. Kaur P, Kallakury BS, Sheehan CE, et al. Survivin and Bcl-2 expression in prostatic adenocarcinomas. Arch Pathol Lab Med. 2004 Jan;128(1):39-43 .
22. Ku JH, Kwak C, Lee HS, et al. Expression of survivin, a novel inhibitor of apoptosis, in superficial transitional cell carcinoma of the bladder. J Urol. 2004 Feb;171(2 Pt 1):631-635.
23. Grabowski P, Kuhnel T, Muhr-Wilkenshoff F, et al. Prognostic value of nuclear survivin expression in oesophageal squamous cell
carcinoma. Br J Cancer. 2003 Jan 13;88(1):115-119.
24. Sarela AI, Macadam RC, Farmery SM, et al . Expression of the antiapoptosis gene, survivin, predicts death from recurrent colorectal carcinoma. Gut. 2000 May;46(5):645-650.
25. Grossman D, McNiFF JM, Li F, et al. Expression of the apoptosis inhibitor, survivin，in nonmelanoma skin cancer and targeting in a keratincyte cell line . Lab Invest , 1999,79:1121-1126.
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