CAJ | 학술논문

背景：一些实验在模型鼠移植瘤中葡萄糖调节蛋白94被敲除后，细胞黏附中断，刺激肝起源细胞增殖，进而促进肝肿瘤的发生，推测葡萄糖调节蛋白94在肝癌中起到保护作用。 　　目的：应用小干扰RNA技术抑制裸鼠卵巢癌模型皮下移植瘤内质网分子伴侣葡萄糖调节蛋白94的基因表达，探讨移植瘤中葡萄糖调节蛋白94基因及蛋白表达的变化对移植瘤生长的影响。 　　方法：从GeneBank中选取人类葡萄糖调节蛋白94基因序列，构建受控于人RNA聚合酶Ⅲ启动子U6的真核表达载体psiSTRIKETM/葡萄糖调节蛋白94。建立人卵巢癌HO-8910细胞株裸鼠皮下接种模型，并将真核表达载体转染入裸鼠移植瘤体内，观察肿瘤生长情况。卵巢癌裸鼠模型经过不同给药方案干预后分为特异性小干扰RNA组，非特异性小干扰RNA组和生理盐水对照组， RT-PCR和免疫组化SP法检测葡萄糖调节蛋白94 mRNA和蛋白在肿瘤内的表达情况，并观察模型裸鼠的移植瘤生长情况。 　　结果与结论：成功构建RNA干扰质粒载体，所有裸鼠模型均接种成功，5 d后即可见皮下肿瘤形成，14 d左右肿瘤直径达7-10 mm。转染质粒完毕后抑瘤率为65.1%，与非特异性小干扰RNA组和生理盐水对照组比较，差异有显著性意义(P<0.01)。psiSTRIKETM/GRP94治疗后瘤体内葡萄糖调节蛋白94 mRNA和蛋白显著下调(P <0.01)。说明靶向葡萄糖调节蛋白94 mRNA的小干扰RNA可显著抑制人卵巢癌裸鼠移植瘤的生长，其机制可能是诱导葡萄糖调节蛋白94 mRNA和蛋白表达下调所致。
배경：일사실험재모형서이식류중포도당조절단백94피고제후，세포점부중단，자격간기원세포증식，진이촉진간종류적발생，추측포도당조절단백94재간암중기도보호작용。 　　목적：응용소간우RNA기술억제라서란소암모형피하이식류내질망분자반려포도당조절단백94적기인표체，탐토이식류중포도당조절단백94기인급단백표체적변화대이식류생장적영향。 　　방법：종GeneBank중선취인류포도당조절단백94기인서렬，구건수공우인RNA취합매Ⅲ계동자U6적진핵표체재체psiSTRIKETM/포도당조절단백94。건립인란소암HO-8910세포주라서피하접충모형，병장진핵표체재체전염입라서이식류체내，관찰종류생장정황。란소암라서모형경과불동급약방안간예후분위특이성소간우RNA조，비특이성소간우RNA조화생리염수대조조， RT-PCR화면역조화SP법검측포도당조절단백94 mRNA화단백재종류내적표체정황，병관찰모형라서적이식류생장정황。 　　결과여결론：성공구건RNA간우질립재체，소유라서모형균접충성공，5 d후즉가견피하종류형성，14 d좌우종류직경체7-10 mm。전염질립완필후억류솔위65.1%，여비특이성소간우RNA조화생리염수대조조비교，차이유현저성의의(P<0.01)。psiSTRIKETM/GRP94치료후류체내포도당조절단백94 mRNA화단백현저하조(P <0.01)。설명파향포도당조절단백94 mRNA적소간우RNA가현저억제인란소암라서이식류적생장，기궤제가능시유도포도당조절단백94 mRNA화단백표체하조소치。
BACKGROUND:After glucose regulated protein 94 (GRP94) was knockout in model mice of transplanted tumor, cel ular adhesion is terminated, thus stimulating the proliferation of liver-derived cel s and promoting the development of liver cancer. We speculate that GRP94 plays a protective role against liver cancer. OBJECTIVE:To investigate the expression of endoplasmic reticulum molecular chaperone GRP94 mRNA and protein with smal interfering RNA technique in nude mice model of transplantable human ovarian carcinoma, and to explore the effect of GRP94 mRNA and protein expression on the growth of transplanted tumor. METHODS:The gene sequences of human GRP94 were obtained from Gene Bank. psiSTRIKETM/GRP94 was constructed, which is eukaryotic expression vector control ed by the U6 promoter of human RNA polymerase Ⅲ. The transplantable model of human ovarian carcinoma in nude mice was established using human ovarian cancer HO-8910 cel line. The eukaryotic expression vector was transfected into the transplanted tumors in nude mice, and the growth of the tumor was observed. The nude mice models were divided into three groups, specific smal interfering RNA group, non-specific smal interfering RNA group and saline control group. The volumes of the subcutaneous tumor were determined. RT-PCR and immunohistochemistry were used to detect the mRNA and protein expression of GRP94 respectively. RESULTS AND CONCLUSION:The recombinant plasmid of RNA interference specific for GRP94 was successful y constructed. The subcutaneous tumors appeared in al the nude mice 5 days after transplantation. The diameter of subcutaneous tumors was 7-10 mm 14 days after transplantation. The growth of subcutaneous tumors in nude mice with interference specific for GRP94 treatment was significantly decreased as compared with non-specific smal interfering RNA group and control group (P<0.05). The proliferation activity was inhibited by 65.1%. The expression of GRP94 mRNA and protein was significantly down-regulated after treatment of psiSTRIKETM/GRP94 (P<0.01). The transfection of psiSTRIKETM/GRP94 could significantly induce inhibitory effects on the growth of ovarian carcinoma in nude mice, and the underlying mechanism is associated with the down-regulated expression of GRP94 mRNA and protein.