临床肿瘤学杂志
臨床腫瘤學雜誌
림상종류학잡지
CHINESE CLINICAL ONCOLOGY
2014年
2期
97-101
,共5页
陈墅圳%聂玉强%杜艳蕾%徐言%沈桂佳
陳墅圳%聶玉彊%杜豔蕾%徐言%瀋桂佳
진서수%섭옥강%두염뢰%서언%침계가
小干扰RNA%肝癌%细胞分裂周期相关蛋白5%增殖%凋亡
小榦擾RNA%肝癌%細胞分裂週期相關蛋白5%增殖%凋亡
소간우RNA%간암%세포분렬주기상관단백5%증식%조망
Small interfering RNA%Hepatocacinoma%Cell division cycle associated 5%Proliferation%Apoptosis
目的：探讨小干扰RNA（siRNA）靶向抑制细胞分裂周期相关蛋白（CDCA5）基因表达对人肝癌HepG2细胞株增殖及凋亡的影响。方法优化siRNA转染条件，将条靶向抑制CDCA5基因的siRNA载体片段（序列、2和）分别高效转染人肝癌HepG2细胞（ A、B和C组），并设阴性对照组（ NC组）及空白组。免疫印迹法检测转染h各组CDCA5蛋白的表达水平。 CCK-8法检测转染、48、72、96、120h各组细胞的增殖能力，Annexin V-FITC/PI 双标记流式细胞术检测各组转染h的凋亡情况。结果 A、B和C组的CDCA5蛋白水平均低于NC组和空白组（ P＜0.05），其中B组的表达率最低，抑制率最高，达.3％，故选择序列进行后续实验。自转染h起，B组的相对增殖率均低于NC组和空白组（ P＜0.05）；B组转染h的相对增殖率为（66.58±2.58）％，均低于其他观察时间点（P＜0.05）。 B组转染h的早期和晚期凋亡率分别为（17.43±2.31）％和（22.37±2.21）％，均高于NC组和空白组（P＜0.05）。结论通过siRNA能够降低HepG2细胞的CDCA5表达水平，有效抑制HepG2细胞的增殖并促进其凋亡。
目的：探討小榦擾RNA（siRNA）靶嚮抑製細胞分裂週期相關蛋白（CDCA5）基因錶達對人肝癌HepG2細胞株增殖及凋亡的影響。方法優化siRNA轉染條件，將條靶嚮抑製CDCA5基因的siRNA載體片段（序列、2和）分彆高效轉染人肝癌HepG2細胞（ A、B和C組），併設陰性對照組（ NC組）及空白組。免疫印跡法檢測轉染h各組CDCA5蛋白的錶達水平。 CCK-8法檢測轉染、48、72、96、120h各組細胞的增殖能力，Annexin V-FITC/PI 雙標記流式細胞術檢測各組轉染h的凋亡情況。結果 A、B和C組的CDCA5蛋白水平均低于NC組和空白組（ P＜0.05），其中B組的錶達率最低，抑製率最高，達.3％，故選擇序列進行後續實驗。自轉染h起，B組的相對增殖率均低于NC組和空白組（ P＜0.05）；B組轉染h的相對增殖率為（66.58±2.58）％，均低于其他觀察時間點（P＜0.05）。 B組轉染h的早期和晚期凋亡率分彆為（17.43±2.31）％和（22.37±2.21）％，均高于NC組和空白組（P＜0.05）。結論通過siRNA能夠降低HepG2細胞的CDCA5錶達水平，有效抑製HepG2細胞的增殖併促進其凋亡。
목적：탐토소간우RNA（siRNA）파향억제세포분렬주기상관단백（CDCA5）기인표체대인간암HepG2세포주증식급조망적영향。방법우화siRNA전염조건，장조파향억제CDCA5기인적siRNA재체편단（서렬、2화）분별고효전염인간암HepG2세포（ A、B화C조），병설음성대조조（ NC조）급공백조。면역인적법검측전염h각조CDCA5단백적표체수평。 CCK-8법검측전염、48、72、96、120h각조세포적증식능력，Annexin V-FITC/PI 쌍표기류식세포술검측각조전염h적조망정황。결과 A、B화C조적CDCA5단백수평균저우NC조화공백조（ P＜0.05），기중B조적표체솔최저，억제솔최고，체.3％，고선택서렬진행후속실험。자전염h기，B조적상대증식솔균저우NC조화공백조（ P＜0.05）；B조전염h적상대증식솔위（66.58±2.58）％，균저우기타관찰시간점（P＜0.05）。 B조전염h적조기화만기조망솔분별위（17.43±2.31）％화（22.37±2.21）％，균고우NC조화공백조（P＜0.05）。결론통과siRNA능구강저HepG2세포적CDCA5표체수평，유효억제HepG2세포적증식병촉진기조망。
Objective To investigate influence of CDCA5 small interfering RNA( CDCA5-siRNA) on proliferation and apop-tosis of human hepatocarcinoma cell line HepG2. Methods The HepG2 cells were divided into siRNA groups ( A, B and C groups) , universal scrambled negative control siRNA group ( NC group) and Blank group. Three CDCA5-siRNAs with different sequences were transfected into HepG2 cells respectively. Western blotting method was used to measure expressions of CDCA5 protein levels at 72h af-ter transfection. The cell proliferation was assessed by CCK-8 at 24, 48, 72, 96 and 120h after transfection. The Annexin V-FITC/PI double-labeled flow cytometry was employed to measure the apoptosis at 48h after transfection. Results The expressions of CDCA5 protein significantly decreased in the CDCA5-siRNA groups ( A,B and C groups) compared with NC group and Blank group( P<0.05) . Group B had the lowest expression rate and highest inhibition rate( 89.3%) . The relative proliferation rate of group B was significantly lower than those of NC group and Blank group since 48 hours after transfection( P<0.05) , and the lowest proliferation rate was ( 66.58 ±2.58)% at 72h in group B. The early and late apoptosis rates were (17.43±2.31) % and (22.37±2.21) % in B group, higher than those of other two groups( P<0.05) . Conclusion siRNA down-regulating the expression of CDCA5 gene in human hepatocarcino-ma cell line HepG2 can inhibit cell proliferation and increase cell apoptosis.
