抗癌药物力达霉素的辅基蛋白对肿瘤组织细胞的结合作用

Study on Binding of Apoprotein of Lidamycin to Cancer Cells

目的研究力达霉素辅基蛋白(idamycin apoprotein,LDP)对肿瘤细胞是否有结合作用,并考察这种结合作用与肿瘤有关生物学指标的相关性。方法用异硫氰酸荧光素酯(fluorescein isothiocyanate,FITC)与LDP进行偶联,以荧光显微镜观察此偶联物对临床常见肿瘤的荧光染色情况,按荧光强弱程度将着色结果分成阳性、可疑、阴性3级。以免疫组化法观察肿瘤生物学指标如ER,P53,Bcl-2表达情况,并将结果也分成阳性、可疑、阴性3级。以Kappa一致性检验和Spearman等级相关性检验判断相关性。结果仅部分乳腺癌显示阳性荧光着色:3例乳腺癌冰冻标本中,有2例显示阳性荧光着色;20例确诊的乳腺癌石蜡标本中,荧光染色显示阳性的11例,可疑的4例,阴性的5例。FITC-LDP偶联物对乳腺癌石蜡标本染色结果与ER、P53和Bcl-2的免疫组化染色结果分别经Kappa一致性检验,均有显著意义(P<0.05);与ER染色结果、P53染色结果和Bcl-2染色结果分别经Spearman等级相关性检验,分别呈负相关(P<0.05)、正相关(P<0.05)和无相关(P>0.05)。结论LDP对乳腺癌细胞存在特异的结合作用,这种结合作用与乳腺癌组织上的ER和突变型P53蛋白分别可能存在负相关性和正相关性。

OBJECTIVE This experiment was set to study binding of the apoprotein to cancer cells and examine the correlation between the binding action and cancer biological index such as ER,P53,Bcl-2. METHODS LDP was linked to FITC, a fluorescent reagent. The FITC-LDP conjugate was incubated with cancer tissue specimens at a certain condition. Fluorescent microscope was used to observe fluorescent intensity of specimens. The fluorescent intensity were graded into 3 levels which included positive, doubtful and negative. The expression of cancer biological index was observed by the method of immunohistochemistry, the staining of which were also graded into 3 levels which contained positive, doubtful and negative. Kappa test or Spearman test were used to analyze the above correlation. RESULTS Only a fraction of breast cancer tissue specimen displayed positive fluorescent staining caused by FITC-LDP conjugate. 2 out of 3 iced specimens showed positive fluorescent staining. 11,4 or 5 out of 20 wax specimens showed positive, doubtful and negative fluorescent staining, respectively. The fluorescent staining and ER, P53 or Bcl-2 immunohistochemistry staining had respective concordance with Kappa test （ P 〈0. 05 ）. The fluorescent staining and ER, P53 or Bcl-2 immunohistochemistry staining showed negative rank correlation, positive rank correlation and no rank correlation respectively with Spearman test （ P 〈 0.05 ）. CONCLUSION LDP may have binding action on breast cancer cells, which might exist negative or positive correlation with ER or mutant P53 protein of breast cancer, respectively.