CAJ | 학술논문

目的：探讨抑制膜联蛋白A7(ANXA7)表达对人肝癌 HepG-2细胞半乳糖凝集素3(galectin-3)的影响以及联合佛波酯(PMA)激活PKC后对 ANXA7表达的影响。方法将细胞分为 siRNA 干扰组、阴性对照组和空白对照组。采用RNA干扰技术将靶向 ANXA7的 siRNA和阴性对照 siRNA脂质体转染法分别转染肝癌 HepG-2细胞,空白对照组不予任何处理。转染48h后,采用 Western blot 和 RT-qPCR法进行抑制效果的鉴定。Western blot和 RT-qPCR法分别检测 ANXA7表达抑制的 HepG-2细胞中galectin-3的表达。转染48h后用100ng/ml PMA处理24h,Western blot法检测ANXA7及PKC的表达。结果靶向 ANXA7的 siRNA可显著抑制 ANXA7的表达；ANXA7表达受抑后的 HepG-2细胞中 galectin-3的表达下降,与阴性对照组和空白对照组相比,差异有显著性(P<0.05)。PMA 处理24h 后,ANXA7及 PKC 表达变化不显著。结论 ANXA7表达抑制的 HepG-2细胞中galectin-3的表达显著下调,这可能是ANXA7表达受抑的细胞行为学发生改变的机制之一。
목적：탐토억제막련단백A7(ANXA7)표체대인간암 HepG-2세포반유당응집소3(galectin-3)적영향이급연합불파지(PMA)격활PKC후대 ANXA7표체적영향。방법장세포분위 siRNA 간우조、음성대조조화공백대조조。채용RNA간우기술장파향 ANXA7적 siRNA화음성대조 siRNA지질체전염법분별전염간암 HepG-2세포,공백대조조불여임하처리。전염48h후,채용 Western blot 화 RT-qPCR법진행억제효과적감정。Western blot화 RT-qPCR법분별검측 ANXA7표체억제적 HepG-2세포중galectin-3적표체。전염48h후용100ng/ml PMA처리24h,Western blot법검측ANXA7급PKC적표체。결과파향 ANXA7적 siRNA가현저억제 ANXA7적표체；ANXA7표체수억후적 HepG-2세포중 galectin-3적표체하강,여음성대조조화공백대조조상비,차이유현저성(P<0.05)。PMA 처리24h 후,ANXA7급 PKC 표체변화불현저。결론 ANXA7표체억제적 HepG-2세포중galectin-3적표체현저하조,저가능시ANXA7표체수억적세포행위학발생개변적궤제지일。
Obj ective To detect the expression of galectin-3 and PKC activated by PMA in annexin A7(ANXA7)knockdown HepG-2 cells.Methods The HepG-2 cells were grouped into experiment group, negative group and blank group.RNA interference technology was usedto transfect ANXA7-targeted siR-NA,and scrambled siRNA into HepG-2 cells with lipofectineTM2000,without any treatment to the blank group.About 48 h after transfection,western blot and RT-qPCR were used to assure the suppression of ANXA7.Then the expression of galectin-3 was detected respectively with western blot and RT-qPCR a-bout 48 h after transfection and about 24 h after 100ng/ml PMA treatment,western blot was used to de-tect the expression of PKC and ANXA7.Results The expression of ANXA7 was significantly suppressed in the cells transfected with ANXA7 siRNA.When ANXA7 expression was suppressed,the expression of galectin-3 decreased significantly compared with that of the negtive control group and blank control group (P<0.05).There was no obvious change in the expression of ANXA7 and PKC about 24 h after 100ng/mL PMA treatment.Conclusion The expression of galectin-3 was down-regulated in the ANXA7 sup-pressed HepG-2 cells,which may be one of the mechanisms for the abnormal behaviour of the ANXA7 knockdown carcinoma cells.