宫颈癌细胞系中HPV 感染与GRP78、JNK 及 CHOP 表达的相关性分析

Correlations of HPV infection with GRP78, JNK and CHOP expressions in cervical cancer cell lines

摘要:

目的 探讨宫颈癌细胞系中HPV（HPV）感染状态与内质网应激相关蛋白表达之间的相关性。 方法 采用免疫组织化学法、Western blot、逆转录聚合酶链反应（RT-PCR）分别检测GRP78、JNK 及 CHOP 蛋白和mRNA 在HPV16（+）Siha、HPV18（+）Hela 及HPV（-）C33a 3 种宫颈癌细胞系中的表达 水平，并采用统计学分析其表达的差异性。结果 免疫组织化学法结果显示，GRP78 蛋白在高危型HPV16（+） Siha 和HPV18（+）Hela 的表达高于HPV（-）C33a（P <0.05），内质网应激相关蛋白JNK、CHOP 在HPV16 （+）Siha 和HPV18（+）Hela 的表达高于HPV（-）C33a（P <0.05）。RT-PCR 及Western blot 结果显示， GRP78、JNK 及CHOP mRNA 及蛋白表达在HPV16（+）Siha 和HPV18（+）Hela 细胞株中高于HPV（-） C33a 细胞株（P <0.05）。结论 宫颈癌中GRP78 和内质网应激相关蛋白JNK、CHOP 的表达与HPV 亚型相关， 内质网应激相关蛋白可能参与了宫颈癌的发生与发展。

Abstract:

Objective To investigate the correlations of HPV infection with the expressions of endoplasmic reticulum stress related proteins in cervical cancer cell lines. Methods Immunocytochemistry, Western blot and reverse transcription polymerase chain reaction (RT-PCR) were employed to detect the protein and mRNA expression levels of glucose-regulated protein 78 (GRP78), C-Jun NH2-terminal kinase (JNK) and CCAAT/enhancer-binding protein homologous protein (CHOP) in HPV16-positive SiHa, HPV18-positive Hela and HPV-negative C33a cervical cancer cell lines. Statistical methods were applied to analyze the statistical differences of their expressions. Results The results of immunocytochemistry showed that GRP78 protein levels of high-risk HPV16-positive SiHa and HPV18-positive Hela cells were significantly higher than that of HPV-negative C33a (P < 0.05), and endoplasmic reticulum stress related proteins JNK and CHOP also had notably higher expressions in high-risk HPV16-positive SiHa and HPV18-positive Hela cells than in the HPV-negative C33a cells (P < 0.05). RT-PCR and Western blot results showed that the mRNA levels of GRP78, JNK and CHOP in HPV16-positive SiHa cells and HPV18-positive Hela cells were significantly higher than those in HPV-negative C33a cells (P < 0.05). Conclusions HPV subtypes are strongly associated with the expressions of GRP78 and endoplasmic reticulum stress related proteins JNK and CHOP, and endoplasmic reticulum stress related proteins may be involved in the occurrence and development of cervical cancer.