组织蛋白表达水平检测方法的比较分析
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摘要
为了得到一种能够准确、快速检测组织蛋白表达水平的方法,收集了12例结肠癌病人的癌组织及癌旁组织,以RRP15 mRNA的表达水平作为参照,比较免疫组织化学染色法和免疫荧光染色法在检测组织蛋白表达水平方面的优劣.免疫组织荧光染色结果显示,12例患者中有8例患者肿瘤组织的RRP15表达为阳性,4例为阴性,其结果与Real-time PCR定量结果高度一致.免疫组织化学染色结果显示,12例患者中有9例患者肿瘤组织的RRP15表达为阳性,其余3例为阴性,其中6号和11号免疫组织化学检测为阴性,而Real-time PCR定量和免疫组织荧光染色结果均为阳性,即免疫组化检测结果与Real-time PCR定量结果的一致性较差.因此认为,免疫荧光染色法是检测组织中蛋白表达水平的首选方法,免疫组织化学染色法可作为一种辅助方法.
In order to obtain a method for exactly and quickly detecting protein expression level in tissue,the cancer tis-sues and paracancerous tissues of 12 patients with colon cancer were collected, and the immunohistochemical and im-munofluorescent staining results for the protein detection were compared. The expression level of RRP15 m RNA was adopt-edas a reference for comparison. The results of immunofluorescent staining showed that among 12 cases of colon cancer, theRRP15 expressions were positive in eight cases and were negative in four cases,which was highly consistent with theresults of quantitative RT-PCR. However,the results of immunohistochemical staining showed that the RRP15 expressionswere positive in nine cases and were negative in three cases. The 6 th and 11 th cases were demonstrated as negative usingimmunohistochemical staining method, which were positive using immunofluorescent staining method and quantitative RT-PCR. That is, the results of immunohistochemical staining method had poor consistency with that of quantitative RT-PCR.According to these results,the immunofluorescent staining can be the first choice for detecting the protein expressionlevel in tissues and immunohistochemical staining could be used as an auxiliary method.
In order to obtain a method for exactly and quickly detecting protein expression level in tissue,the cancer tis-sues and paracancerous tissues of 12 patients with colon cancer were collected, and the immunohistochemical and im-munofluorescent staining results for the protein detection were compared. The expression level of RRP15 m RNA was adopt-edas a reference for comparison. The results of immunofluorescent staining showed that among 12 cases of colon cancer, theRRP15 expressions were positive in eight cases and were negative in four cases,which was highly consistent with theresults of quantitative RT-PCR. However,the results of immunohistochemical staining showed that the RRP15 expressionswere positive in nine cases and were negative in three cases. The 6 th and 11 th cases were demonstrated as negative usingimmunohistochemical staining method, which were positive using immunofluorescent staining method and quantitative RT-PCR. That is, the results of immunohistochemical staining method had poor consistency with that of quantitative RT-PCR.According to these results,the immunofluorescent staining can be the first choice for detecting the protein expressionlevel in tissues and immunohistochemical staining could be used as an auxiliary method.
引文
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[6] ZHOU B,WU Q,CHEN G,et al. NOP14 promotes proliferation andmetastasis of pancreatic cancer cells[J]. Cancer Letters,2012,322(2):195-203.
[7]杜永星,李宗泽,刘长征,等.胰腺癌小鼠模型中核仁蛋白14和CD31表达与肿瘤进展的相关性研究[J].中华外科杂志,2017,55(6):463-467.DU Y X,LI Z Z,LIU C Z,et al. Expression of nucleolar protein 14and CD31 in pancreatic cancer mouse model and its correlation withtumor progression[J]. Chinese Journal of Surgery,2017,55(6):463-467(in Chinese).
[8]王伟超.核仁蛋白EBP2反馈调控转录因子c-Myc影响r DNA转录、细胞增殖以及肿瘤形成[D].上海:华东师范大学,2015.WANG W C. A Positive Feedback Loop Between EBP2 and c-Myc Reg-ulates r DNA Transcription,Cell Proliferation and Tumorigenesis[D].Shanghai:East China Normal University,2015(in Chinese).
[9] LI J,YU L,ZHANG H Z,et al. Down-regulation of pescadillo inhibitsproliferation and tumorigenicity of breast cancer cells[J]. Cancer Sci-ence,2010,100(12):2255-2260.
[10] CHARPENTIER A H,BEDNAREK A K,DANIEL R L,et al. Effectsof estrogen on global gene expression:Identification of novel targets ofestrogen action[J]. Cancer Research,2000,60(21):5977-5983.
[11] ZHANG J H,YANG Y,WU J R. B23 interacts with PES1 and isinvolved in nucleolar localization of PES1[J]. Acta Biochimica Et Bio-physica Sinica,2009,41(12):991-997.
[12] PARIKH C R,MISHRA J,THIESSEN-PHILBROOK H,et al. UrinaryIL-18 is an early predictive biomarker of acute kidney injury after car-diac surgery[J]. Kidney International,2006,70(1):199-203.
[13] MISHRA J,DENT C,TARABISHI R,et al. Neutrophil gelatinase-associated lipocalin(NGAL)as a biomarker for acute renal injuryafter cardiac surgery[J]. Lancet,2005,365(9466):1231-1238.
[14]施国伟,潘煜海,何家扬. Survivin与泌尿系肿瘤[J].国际泌尿系统杂志,2003,23(2):120-122.SHI G W,PAN Y H,HE J Y. Survivin and urinary tumors[J]. Interna-tional Journal of Urology,2003,23(2):120-122(in Chinese).
[15] ROBERTSON D,SAVAGE K,REISFILHO J S,et al. Multiple immu-nofluorescence labelling of formalin-fixed paraffin-embedded(FFPE)tissue[J]. BMC Cell Biology,2008,9(1):13-23.