摘要
目的 :检测血管内皮生长因子C(VEGF C)mRNA在人乳腺癌细胞株MDA MB 2 31及MCF 7中的定位、定量表达。方法 :根据VEGF C基因序列 ,设计合成地高辛标记的特异性寡核苷酸探针 ,运用原位杂交方法检测培养的细胞株MDA MB 2 31及MCF 7中VEGF CmRNA的定位表达 ;并运用半定量RT PCR方法检测了两种细胞中VEGF C相对于看家基因 β 肌动蛋白 (β actin)的表达水平。结果 :原位杂交检测到MDA MB 2 31及MCF 7细胞的胞浆中有阳性蓝色颗粒 ,而阴性对照细胞的胞浆中则均无阳性颗粒 ;半定量RT PCR检测到两种细胞中VEGF C基因均以较高水平表达 ,相对 β actin的表达水平分别为 0 .89和 0 .6 8。结论 :人乳腺癌细胞株MDA MB 2 31及MCF 7细胞能够以较高水平转录VEGF
Objective: The study was designed to examine the localizations and expression levels of vascular endothelial growth factor C (VEGF-C) mRNA in human breast cancer cell lines MDA-MB-231 and MCF-7. Methods: A′pecific oligonucleotide probe labeled with digoxin was designed and constructed according to the VEGF-C gene sequence,and in situ hybridization was employed to detect the localizations of VEGF-C mRNA in the cultured human breast cancer cell lines MDA-MB-231 and MCF-7. In addition semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was employed to measure the expression levels of VEGF-C mRNA in both cell lines in relation to those of housekeeping gene β-actin. Results: Blue positive staining granules for VEGF-C mRNA were observed in the cytoplasm of these cells by in situ hybridization but no positive staining granules were found in either of the control cells. RT-PCR analysis showed that VEGF-C mRNA was expressed at relatively high levels (0.89 and 0.68) in the two kinds of cells with respect to that of housekeeping gene β-actin. Conclusion: Human breast cancer cell lines MDA-MB-231 and MCF-7 can transcribe VEGF-C mRNA at relatively higher levels.
出处
《山东大学学报（医学版）》
CAS
2002年第3期219-221,共3页
Journal of Shandong University:Health Sciences
