CAJ | 학술논문

目的：使用小干扰RNA（ siRNA）抑制人肝癌SK-Hep-1细胞中蛋白激酶Cε（ PKCε）的表达并对抑制效果进行测定，检测沉默后SK-Hep-1细胞增殖和侵袭能力的变化。方法：培养SK-Hep-1细胞，分为PKCε-siR-NA组、阴性对照（NC-siRNA）组和未行处理（control）组。采用MTT比色法观察各组细胞增殖能力的变化，采用Tr-answell法观察各组细胞侵袭能力的变化，Western blotting观察细胞核增殖抗原Ki67、基质金属蛋白酶9（ MMP-9）等功能蛋白的表达，萤光素酶报告基因检测NF-κB信号通路的活性。结果：与对照组相比， PKCε-siRNA组PKCεmRNA和蛋白表达水平都明显降低（P＜0.01），Ki67和MMP-9蛋白表达水平降低，细胞增殖能力明显降低（P＜0.01），PKCε-siRNA组穿过Transwell膜的细胞少于对照组（P＜0．01），且NF-κB转录活性下降（P＜0．01）。结论：通过siRNA技术降低PKCε表达可抑制肝癌SK-Hep-1细胞增殖及其侵袭能力，其抑制作用可能与抑制NF-κB通路的转录活性有关。
목적：사용소간우RNA（ siRNA）억제인간암SK-Hep-1세포중단백격매Cε（ PKCε）적표체병대억제효과진행측정，검측침묵후SK-Hep-1세포증식화침습능력적변화。방법：배양SK-Hep-1세포，분위PKCε-siR-NA조、음성대조（NC-siRNA）조화미행처리（control）조。채용MTT비색법관찰각조세포증식능력적변화，채용Tr-answell법관찰각조세포침습능력적변화，Western blotting관찰세포핵증식항원Ki67、기질금속단백매9（ MMP-9）등공능단백적표체，형광소매보고기인검측NF-κB신호통로적활성。결과：여대조조상비， PKCε-siRNA조PKCεmRNA화단백표체수평도명현강저（P＜0.01），Ki67화MMP-9단백표체수평강저，세포증식능력명현강저（P＜0.01），PKCε-siRNA조천과Transwell막적세포소우대조조（P＜0．01），차NF-κB전록활성하강（P＜0．01）。결론：통과siRNA기술강저PKCε표체가억제간암SK-Hep-1세포증식급기침습능력，기억제작용가능여억제NF-κB통로적전록활성유관。
AIM:To investigate the inhibitory effect of small interfering RNA ( siRNA) on the expression of protein kinase Cε( PKCε) in human hepatoma SK-Hep-1 cells, and the biological behaviors of the transduced cells , inclu-ding proliferation and invasion , were investigated.METHODS:The cultured SK-Hep-1 cells were divided into 3 groups, including PKCε-siRNA group , negative control ( NC)-siRNA group and control group .MTT assay was used to analyze the proliferation of the SK-Hep-1 cells in the respective groups , while invasion potency was determined by Transwell assay .The protein levels of functional biomarkers such as Ki 67 and matrix metalloproteinase 9 ( MMP-9 ) were measured by Western blotting .The Luciferase reporter gene assay was used to explore the activity of the NF-κB pathway .RESULTS:PKCεex-pression in SK-Hep-1 cells transfected with PKCε-siRNA was significantly down-regulated at both mRNA and protein levels compared with that in the normal SK-Hep-1 cells (P<0.01), with the decreases in the protein levels of Ki67 and MMP-9. The invasion and proliferation of SK-Hep-1 cells were obviously inhibited in PKCε-siRNA group compared with control group (P<0.01).Furthermore, the transcriptional activity of NF-κB was down-regulated when PKCε was effectively in-hibited by PKCε-siRNA (P<0.01).CONCLUSION:Down-regulation of PKCεinhibits the proliferation and invasion of hepatic carcinoma cells , which might be mediated via the NF-κB signaling pathway .