南通大学学报（医学版）
南通大學學報（醫學版）
남통대학학보（의학판）
JOURNAL OF NANTONG UNIVERSITY(MEDICAL SCIENCES)
2015年
1期
4-8
,共5页
陈昱延%刘国梁%魏郦娴%蒋大伟%沈天怡%徐文浩%倪润洲
陳昱延%劉國樑%魏酈嫻%蔣大偉%瀋天怡%徐文浩%倪潤洲
진욱연%류국량%위역한%장대위%침천이%서문호%예윤주
肝细胞肝癌%小谷氨酰胺三角四肽重复区蛋白α抗体%增殖%细胞周期
肝細胞肝癌%小穀氨酰胺三角四肽重複區蛋白α抗體%增殖%細胞週期
간세포간암%소곡안선알삼각사태중복구단백α항체%증식%세포주기
hepatocellular carcinoma%small glutamine-rich tetratricopeptide repeat-containing protein alpha%proliferation%cell cycle
目的：研究靶向下调小谷氨酰胺三角四肽重复区蛋白α抗体(small glutamine-rich tetratricopeptide repeat-containing protein alpha, SGTA)的表达对肝癌HuH7细胞周期及细胞增殖的影响。方法：选择HuH7细胞系作为研究对象，进行血清饥饿释放同步化处理后采用Western Blot方法检测SGTA蛋白表达水平变化；通过SGTA特异性siRNA下调HuH7细胞中SGTA的表达，采用细胞计数盒8(cell counting kit 8, CCK8)、流式细胞术等方法检测细胞增殖能力及细胞周期。结果：血清饥饿使HuH7细胞生长周期停滞，血清释放后刺激HuH7细胞增殖，Western Blot显示SGTA﹑增殖细胞核抗原(proliferating cell nuclear antigen, PCNA)蛋白随着细胞周期的进展表达增加，而细胞周期负性调控因子p27kip1蛋白表达却下降。 SGTA缺失可引起cyclin A、cyclin B的表达下降、细胞增殖的下降以及细胞周期的阻滞。结论：SGTA特异性siRNA能明显抑制肝癌HuH7细胞的增殖，细胞周期阻滞，提示其可为肝癌的靶基因治疗提供新的分子靶点。
目的：研究靶嚮下調小穀氨酰胺三角四肽重複區蛋白α抗體(small glutamine-rich tetratricopeptide repeat-containing protein alpha, SGTA)的錶達對肝癌HuH7細胞週期及細胞增殖的影響。方法：選擇HuH7細胞繫作為研究對象，進行血清饑餓釋放同步化處理後採用Western Blot方法檢測SGTA蛋白錶達水平變化；通過SGTA特異性siRNA下調HuH7細胞中SGTA的錶達，採用細胞計數盒8(cell counting kit 8, CCK8)、流式細胞術等方法檢測細胞增殖能力及細胞週期。結果：血清饑餓使HuH7細胞生長週期停滯，血清釋放後刺激HuH7細胞增殖，Western Blot顯示SGTA﹑增殖細胞覈抗原(proliferating cell nuclear antigen, PCNA)蛋白隨著細胞週期的進展錶達增加，而細胞週期負性調控因子p27kip1蛋白錶達卻下降。 SGTA缺失可引起cyclin A、cyclin B的錶達下降、細胞增殖的下降以及細胞週期的阻滯。結論：SGTA特異性siRNA能明顯抑製肝癌HuH7細胞的增殖，細胞週期阻滯，提示其可為肝癌的靶基因治療提供新的分子靶點。
목적：연구파향하조소곡안선알삼각사태중복구단백α항체(small glutamine-rich tetratricopeptide repeat-containing protein alpha, SGTA)적표체대간암HuH7세포주기급세포증식적영향。방법：선택HuH7세포계작위연구대상，진행혈청기아석방동보화처리후채용Western Blot방법검측SGTA단백표체수평변화；통과SGTA특이성siRNA하조HuH7세포중SGTA적표체，채용세포계수합8(cell counting kit 8, CCK8)、류식세포술등방법검측세포증식능력급세포주기。결과：혈청기아사HuH7세포생장주기정체，혈청석방후자격HuH7세포증식，Western Blot현시SGTA﹑증식세포핵항원(proliferating cell nuclear antigen, PCNA)단백수착세포주기적진전표체증가，이세포주기부성조공인자p27kip1단백표체각하강。 SGTA결실가인기cyclin A、cyclin B적표체하강、세포증식적하강이급세포주기적조체。결론：SGTA특이성siRNA능명현억제간암HuH7세포적증식，세포주기조체，제시기가위간암적파기인치료제공신적분자파점。
Objective: To investigate the change of cell cycle and proliferation of hepatic cell carcinoma through targeted <br> down-regulation the expression of small glutamine-rich tetratricopeptide repeat-containing protein alpha(SGTA) by the specific SGTA-siRNA. Methods: HuH7 cells were selected as the research object, Western Blot was used to detect SGTA protein expression level changes after serum starvation and refeeding, and analyze the role of SGTA in cell proliferation. Then, after knockdown of SGTA in HCC cells using small interfering RNAs, cell counting kit-8(CCK8) assay and flow cytometry were adopted respectively to determine cell proliferation and cell cycle progression in the cells. Results: The cell cycle of HuH7 cells was blocked by serum starvation, but the HuH7 cells were reentering the cell cycle after serum addition. Western Blot showed that the expression of SGTA and proliferatry cell nuclear antigen(PCNA) was increased, whereas the expression of p27 kip1 was inversely diminished after serum stimulation in HuH7 cells during cell cycle progression. siRNA analysis showed that SGTA depletion could inhibited cell proliferation and resulted in cell cycle arrest. Conclusions: siRNA-targeted SGTA could inhibit the cell proliferation and cell cycle of HuH7 cells which suggests that SGTA might serve as a new molecular target for the treatment of HCC.
