肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2010年
3期
186-188
,共3页
肺肿瘤%基因%聚合酶链反应
肺腫瘤%基因%聚閤酶鏈反應
폐종류%기인%취합매련반응
Lung neoplasms%Gene%Polymerase chain reaction
目的 分析比较广泛应用的内参基因β-葡萄糖苷酸酶(GUSB)、3-磷酸甘油醛脱氢酶(GAPDH)和β2-微球蛋白(β 2-M)在中国人的肺癌组织及正常肺组织中的表达稳定性.方法 采用实时定量PCR技术(qRT-PCR)检测这3个看家基因在20例肺癌组织及相应癌旁正常肺组织中的表达情况,并用两款国际通用的内参基因筛选软件GeNorm和NormFinder分析其稳定性.结果 GAPDH在组内及组间的表达差异均最大[肿瘤与正常组内标准差(s)分别为1.07和0.93;|△Ct|=2.01±1.06;P=0.000].用两个软件分析均以3个基因的平均值做内参最为稳定,但t检验结果显示以GUSB和β2-M二者平均值作为内参的组内及组间表达差异最小,并且是唯一一个组合在肿瘤和正常组间差异无统计学意义(肿瘤与正常组内s分别为0.53和0.79;|△Ct|=0.73±0.53;P=0.053).结论 3个看家基因均不适于单独作为肺组织荧光定量PCR检测的内参;GUSB和β2-M的均值是这3个基因不同组合中作为肺组织标本内参的最好选择.
目的 分析比較廣汎應用的內參基因β-葡萄糖苷痠酶(GUSB)、3-燐痠甘油醛脫氫酶(GAPDH)和β2-微毬蛋白(β 2-M)在中國人的肺癌組織及正常肺組織中的錶達穩定性.方法 採用實時定量PCR技術(qRT-PCR)檢測這3箇看傢基因在20例肺癌組織及相應癌徬正常肺組織中的錶達情況,併用兩款國際通用的內參基因篩選軟件GeNorm和NormFinder分析其穩定性.結果 GAPDH在組內及組間的錶達差異均最大[腫瘤與正常組內標準差(s)分彆為1.07和0.93;|△Ct|=2.01±1.06;P=0.000].用兩箇軟件分析均以3箇基因的平均值做內參最為穩定,但t檢驗結果顯示以GUSB和β2-M二者平均值作為內參的組內及組間錶達差異最小,併且是唯一一箇組閤在腫瘤和正常組間差異無統計學意義(腫瘤與正常組內s分彆為0.53和0.79;|△Ct|=0.73±0.53;P=0.053).結論 3箇看傢基因均不適于單獨作為肺組織熒光定量PCR檢測的內參;GUSB和β2-M的均值是這3箇基因不同組閤中作為肺組織標本內參的最好選擇.
목적 분석비교엄범응용적내삼기인β-포도당감산매(GUSB)、3-린산감유철탈경매(GAPDH)화β2-미구단백(β 2-M)재중국인적폐암조직급정상폐조직중적표체은정성.방법 채용실시정량PCR기술(qRT-PCR)검측저3개간가기인재20례폐암조직급상응암방정상폐조직중적표체정황,병용량관국제통용적내삼기인사선연건GeNorm화NormFinder분석기은정성.결과 GAPDH재조내급조간적표체차이균최대[종류여정상조내표준차(s)분별위1.07화0.93;|△Ct|=2.01±1.06;P=0.000].용량개연건분석균이3개기인적평균치주내삼최위은정,단t검험결과현시이GUSB화β2-M이자평균치작위내삼적조내급조간표체차이최소,병차시유일일개조합재종류화정상조간차이무통계학의의(종류여정상조내s분별위0.53화0.79;|△Ct|=0.73±0.53;P=0.053).결론 3개간가기인균불괄우단독작위폐조직형광정량PCR검측적내삼;GUSB화β2-M적균치시저3개기인불동조합중작위폐조직표본내삼적최호선택.
Objective To analyze the expression stability of the three widely used reference genes β-glueuronidase (GUSB), glycera]dehydes-3-phosphate dehydrogenase (GAPDH), β2-microglobulin (β2-M) in Chinese lung cancer tissue specimens and normal lung tissue specimens. Methods Gene expression wasmeasured by quantitative real time PCR and expression stability was analyzed with two widely used softwares genorm and normfinder. Results The intra-and inter-group difference of GAPDH is maximum (The intra and inter-group s is 1.07 and 0.93 respectively, |△ Ct|=2.01±1.06; P =0.000). The mean of these three genes' Ct value is the most stable one analyzed by the two softwares. But the t test showed that the mean of Ct value of GUSB and β2-M is the unique combination that had the minimum intra-and inter-group difference, with no statistically significant differences between normal and malignant samples (The intra-and inter-group s is 0.53 and 0.79 respective]y, |△Ct|= 0.73±0.53; P =0.053). Conclusion It is inappropriate to normalize data derived from lung tissue specimens using one of these three housekeeping genes alone. Among the different combinations of these three genes, the mean of the Ct values of GUSB and β2-M is the best choice as the internal control of lung tissue specimens.
