中国药理学通报2016，Vol.32Issue(11)：1510-1515,1516,7.DOI:10.3969/j.issn.1001-1978.2016.11.007
缝隙连接蛋白43对人非小细胞肺癌HCC827细胞吉非替尼获得性耐药机制的初步研究
Effect of Cx43 on acquired gefitinib-resistance mechanisms in human NSCLC HCC827 cells
摘要
Abstract
Aim To explore the effect of connexin 43 ( Cx 4 3 ) on acquired gefitinib-resistance in human non small cell lung cancer ( NSCLC ) . Methods HCC827 GR, a gefitinib-resistant ( GR) NSCLC cell lines from their parental cells was established by gradually in-creasing the concentration of gefitinib. Gefitinib effica-cy in HCC827 and HCC827 GR cells was detected by MTT assay. Expression of Cx43 mRNA in HCC827 and HCC827 GR cells was determined by RT-PCR. The protein expressions of Cx43 and phospho-Akt ( p-Akt) in these cells were detected by Western blot. The func-tional gap junction intercellular communication ( GJIC ) was measured by parachute assay. The cellular locali-zation of Cx43 protein was evaluated by immunofluores-cence staining. Results MTT assay showed less ge-fitinib cytotoxicity in HCC827 GR cells than that in their parental cells with IC50 of (10. 84 ± 0. 021) μmol ·L-1 versus (0. 07 ± 0. 019) μmol·L-1 , respective-ly. Moreover, both mRNA and protein expressions of Cx43 in HCC827 GR cells were significantly lower than those in HCC827 cells ( P<0. 05 ) . However, the p-Akt protein in HCC827 GR cells was obviously higher than that in HCC827 cells ( P<0. 05 ) . Furthermore, treatment with LY294002 caused a significant reduced p-Akt expression, but a significant increased Cx43 ex-pression in HCC827 GR cells. Moreover, no detecta-ble GJIC was found in HCC827 and their GR cells with or without RA ( a well-defined GJIC enhancer ) treat-ment. Immunofluorescence staining clearly showed that Cx43 protein accumulated in the cytoplasm of HCC827 and their GR cells. Conclusion The down-regulation of Cx43 expression in cytoplasm of HCC827 GR cells may contribute to the acquired gefitinib resistance in NSCLC cells by GJIC-independent activation of PI3 K/Akt signaling pathway.关键词
非小细胞肺癌/缝隙连接蛋白43/吉非替尼/获得性耐药/PI3K/Akt/缝隙连接功能Key words
NSCLC/Cx43/gefitinib/acquired resist-ance/PI3 K/Akt signaling pathway/GJIC分类
医药卫生引用本文复制引用
骆敏,罗燕妹,覃贵慧,滕翠芳,王翰林,阳洁..缝隙连接蛋白43对人非小细胞肺癌HCC827细胞吉非替尼获得性耐药机制的初步研究[J].中国药理学通报,2016,32(11):1510-1515,1516,7.基金项目
国家自然科学基金资助项目( No 81260324) （ No 81260324）
教育部高等学校博士学科点专项科研基金新教师类资助项目( No 20124503120008) （ No 20124503120008）
