摘要
目的观察甲基化结合域蛋白1（MBD1）和葡萄糖调节蛋白78（GRP78）在胰腺癌侵袭转移中的作用。方法检测毒胡萝卜素（Tg）处理后的BxPC-3细胞GRP78的表达及细胞迁移能力的变化；对MBD1-siRNA转染后的BxPC-3细胞MBD1和GRP78的表达、细胞侵袭能力的变化进行分析；免疫组织化学法检测40例胰腺癌组织和15例正常胰腺组织中MBD1和GRP78的表达。结果Tg处理BxPC-3细胞后，GRP78表达明显上升，细胞的迁移能力增加；MBD1-siRNA转染Bxpc-3细胞后，MBD1和GRP78的表达同时下降，细胞侵袭力减弱；MBD1和GRP78在胰腺癌中的表达明显高于正常胰腺组织（P〈0．05），两者的阳性表达呈正相关（r=0．594，P〈0．01），并均与淋巴结转移有关（P〈0．05）。结论GRP78可能受到MBD1的甲基化调控，并与胰腺癌的侵袭转移有关。
Objective To investigate the role of methyi-CpG binding domain protein 1 （MBD1） and glucose regulated protein 78 （ GRP78 ） in the invasion and metastasis of pancreatic cancer and their interaction. Methods After treatment of pancreatic cancer cell line BxPC-3 with thapsigargin （Tg）, quantitative reverse transcription-polymerase chain reaction （RT-PCR） and Western blotting were used to detect the expression of GRP78 mRNA and protein. By using wound healing test, the changes in migration were evaluated. By using RNA interference technique, the MBDI-siRNA plasmid was transfected into BxPC-3 cells, quantitative RT-PCR and Western blotting were used to analyze MBD1 and GRP78 mRNA and protein expression after transfection, and Transwell invasion assay was used to measure cell migration. Immunohistochemically, the expression of MBD1 and GRP78 proteins was detected in 40 cases of pancreatic cancer and 15 cases of normal pancreatic tissue. Results After Tg treatment of BxPC-3 cells （Tg/BxPC-3）, GRP78 mRNA and protein ex- pression was significantly increased, and the number of migratory BxPC-3 was also increased. After MBD1- siRNA plasmid was transfected into Tg/BxPC-3, MBD1 and GRP78 mRNA and protein expression was simul- taneously decreased, while the invasion ability of BxPC-3 diminished. The positive rate of MBD1 and GRP78 was 70. 0% （28） and 67.5% （26） respectively out of 40 cases of pancreatic cancer specimens, significantly higher than that of the normal pancreatic tissue （P 〈0. 05）. There was a positive correlation between MBD1 and GRP78 protein expression （r = 0. 594, P 〈 0. 01 ）, and both MBD1 and GRP78 expression was associated with lymph node metastasis （P 〈 0. 05）. Conclusion Endoplasmic reticulum molecular chaperone GRP78 may act as an important target of MBD1 via methylation, which may have an impact on the characteristics of pancreatic cancer invasion and metastasis. The regulation of GRP78 expression by MBD1 may change the nature of pancreatic cancer invasion a
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第7期1335-1338,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（81001058）
高等学校博士学科点专项科研基金资助项目（20100071120092）
关键词
胰腺癌
内质网应激
葡萄糖调节蛋白
Pancreatic carcinoma
Endoplasmic retieulum stress
Glucose regulated protein
