中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2014年
7期
499-502
,共4页
刘曦霖%简强%苗叶%黄敏%李承新
劉晞霖%簡彊%苗葉%黃敏%李承新
류희림%간강%묘협%황민%리승신
Rab23%慢病毒感染%癌,鳞状细胞%细胞系,肿瘤%细胞增殖
Rab23%慢病毒感染%癌,鱗狀細胞%細胞繫,腫瘤%細胞增殖
Rab23%만병독감염%암,린상세포%세포계,종류%세포증식
Rab23%Lentivirus infections%Carcinoma,squamous cell%Cell line,tumor%Cell proliferation
目的 观察Rab23分子对鳞状细胞癌(简称鳞癌)细胞Sa3增殖的影响及机制.方法 将鳞状细胞癌Sa3细胞分为4组:正常对照组[转染绿色荧光蛋白(eGFP)]、Rab23过表达组(转染eGFP标记的Rab23过表达质粒)、Rab23沉默组(转染Rab23 shRNA质粒)、Rab23空载体组(转染空载体).平板克隆形成实验、流式细胞仪检测Rab23过表达和沉默对Sa3细胞增殖能力的影响.Western印迹检测Rab23过表达和沉默对Sa3细胞Erk/p-Erk表达水平的变化.两组间均数比较采用t检验,多组间均数比较采用单因素方差分析,多组资料两两比较使用Bonferroni's多重比较分析.结果 通过构建质粒、慢病毒感染,获得Rab23过表达和Rab23沉默的稳定Sa3细胞株.平板克隆形成实验结果显示,Rab23过表达组克隆形成率为2.3％±0.2％,与正常对照组(3.6％±0.3％)相比,Sa3细胞增殖能力降低(P＜0.05),而Rab23沉默组克隆形成率(4.1％±0.2％)较空载体组(1.8％±0.0％)增殖能力明显提高(P＜0.01).细胞周期检测显示,Rab23过表达可引起Sa3细胞G1期阻滞,Rab23过表达组增殖指数(0.581±0.035)较正常对照组(0.698±0.018)降低(P＜0.05),而Rab23沉默组增殖指数(0.567±0.015)较空载体组(0.444±0.014)明显提高(P＜0.01).Western印迹结果显示,与正常对照组相比,Rab23过表达组和Rab23沉默组Sa3细胞Erk表达水平无变化,但Rab23过表达组磷酸化Erk表达水平较正常对照组降低,Rab23沉默组磷酸化Erk表达水平较空载体组升高.结论 Rab23对鳞癌Sa3细胞增殖起抑制作用,这种抑制作用可能与Erk通路有关.
目的 觀察Rab23分子對鱗狀細胞癌(簡稱鱗癌)細胞Sa3增殖的影響及機製.方法 將鱗狀細胞癌Sa3細胞分為4組:正常對照組[轉染綠色熒光蛋白(eGFP)]、Rab23過錶達組(轉染eGFP標記的Rab23過錶達質粒)、Rab23沉默組(轉染Rab23 shRNA質粒)、Rab23空載體組(轉染空載體).平闆剋隆形成實驗、流式細胞儀檢測Rab23過錶達和沉默對Sa3細胞增殖能力的影響.Western印跡檢測Rab23過錶達和沉默對Sa3細胞Erk/p-Erk錶達水平的變化.兩組間均數比較採用t檢驗,多組間均數比較採用單因素方差分析,多組資料兩兩比較使用Bonferroni's多重比較分析.結果 通過構建質粒、慢病毒感染,穫得Rab23過錶達和Rab23沉默的穩定Sa3細胞株.平闆剋隆形成實驗結果顯示,Rab23過錶達組剋隆形成率為2.3％±0.2％,與正常對照組(3.6％±0.3％)相比,Sa3細胞增殖能力降低(P＜0.05),而Rab23沉默組剋隆形成率(4.1％±0.2％)較空載體組(1.8％±0.0％)增殖能力明顯提高(P＜0.01).細胞週期檢測顯示,Rab23過錶達可引起Sa3細胞G1期阻滯,Rab23過錶達組增殖指數(0.581±0.035)較正常對照組(0.698±0.018)降低(P＜0.05),而Rab23沉默組增殖指數(0.567±0.015)較空載體組(0.444±0.014)明顯提高(P＜0.01).Western印跡結果顯示,與正常對照組相比,Rab23過錶達組和Rab23沉默組Sa3細胞Erk錶達水平無變化,但Rab23過錶達組燐痠化Erk錶達水平較正常對照組降低,Rab23沉默組燐痠化Erk錶達水平較空載體組升高.結論 Rab23對鱗癌Sa3細胞增殖起抑製作用,這種抑製作用可能與Erk通路有關.
목적 관찰Rab23분자대린상세포암(간칭린암)세포Sa3증식적영향급궤제.방법 장린상세포암Sa3세포분위4조:정상대조조[전염록색형광단백(eGFP)]、Rab23과표체조(전염eGFP표기적Rab23과표체질립)、Rab23침묵조(전염Rab23 shRNA질립)、Rab23공재체조(전염공재체).평판극륭형성실험、류식세포의검측Rab23과표체화침묵대Sa3세포증식능력적영향.Western인적검측Rab23과표체화침묵대Sa3세포Erk/p-Erk표체수평적변화.량조간균수비교채용t검험,다조간균수비교채용단인소방차분석,다조자료량량비교사용Bonferroni's다중비교분석.결과 통과구건질립、만병독감염,획득Rab23과표체화Rab23침묵적은정Sa3세포주.평판극륭형성실험결과현시,Rab23과표체조극륭형성솔위2.3％±0.2％,여정상대조조(3.6％±0.3％)상비,Sa3세포증식능력강저(P＜0.05),이Rab23침묵조극륭형성솔(4.1％±0.2％)교공재체조(1.8％±0.0％)증식능력명현제고(P＜0.01).세포주기검측현시,Rab23과표체가인기Sa3세포G1기조체,Rab23과표체조증식지수(0.581±0.035)교정상대조조(0.698±0.018)강저(P＜0.05),이Rab23침묵조증식지수(0.567±0.015)교공재체조(0.444±0.014)명현제고(P＜0.01).Western인적결과현시,여정상대조조상비,Rab23과표체조화Rab23침묵조Sa3세포Erk표체수평무변화,단Rab23과표체조린산화Erk표체수평교정상대조조강저,Rab23침묵조린산화Erk표체수평교공재체조승고.결론 Rab23대린암Sa3세포증식기억제작용,저충억제작용가능여Erk통로유관.
Objective To evaluate the effect of Rab23 on the proliferation of a squamous cell carcinoma cell line Sa3,and to investigate its mechanisms.Methods Cultured Sa3 cells were classified into four groups:normal control group transfected with green fluorescent protein (GFP),Rab23-overexpressing group transfected with a GFP-labelled Rab23-overexpressing plasmid,Rab23-silencing group transfected with a plasmid carrying a Rab23-targeting shRNA,empty vector group transfected with an empty vector.After additional culture for different durations,plate colony formation assay and flow cytometry were performed to evaluate the proliferative activity of Sa3 cells,and Western blot was conducted to detect the expression of Erl/phosphorylated-Erk in Sa3 cells.Statistical analysis was carried out by t test,one-way analysis of variance and Bonferroni's multiple comparison test.Results Stable Sa3 cell lines with overexpression or silencing of Rab23 were established by plasmid construction and lentivirus-mediated transfection.The plate colony formation assay showed that the colony formation rate was significantly lower in the Rab23-overexpressing group than in the normal control group (2.3％ ± 0.2％ vs.3.6％ ± 0.3％,P ＜ 0.05),but higher in the Rab23-silencing group than in the empty vector group (4.1％ ± 0.2％ vs.1.8％ ± 0.03％,P ＜ 0.01).Rab23 overexpression induced G1 phase arrest in Sa3 cells.The proliferation index was significantly decreased in the Rab23-overexpressing group compared with the normal control group (0.581 ± 0.035 vs.0.698 ± 0.018,P ＜ 0.05),but increased in the Rab23-silencing group compared with the empty vector group (0.567 ± 0.015 vs.0.444 ± 0.014,P ＜ 0.01).As Western blot showed,there were no significant changes in the expression of Erk in the Rab23-silencing or-overexpressing group compared with the normal control group,whereas the expression of p-Erk was attenuated in the Rab23-overexpressing group compared with the normal control group,but enhanced in the Rab23-silencing group compared with the empty vector group.Conclusions Rab23 could inhibit the proliferation of Sa3 cells,which may be associated with the Erk pathway.
