中华放射医学与防护杂志
中華放射醫學與防護雜誌
중화방사의학여방호잡지
Chinese Journal of Radiological Medicine and Protection
2014年
11期
813-816,866
,共5页
周美娟%黄海波%林志祥%丁振华
週美娟%黃海波%林誌祥%丁振華
주미연%황해파%림지상%정진화
miR-365%紫外线B%皮肤鳞状细胞癌%癌基因
miR-365%紫外線B%皮膚鱗狀細胞癌%癌基因
miR-365%자외선B%피부린상세포암%암기인
MiR-365%UVB%Cutaneous squamous cell carcinoma%Oncogene
目的 探讨紫外线(UVB)辐射敏感miR-365在皮肤鳞状细胞癌发生中的作用.方法 取对数生长期正常人皮肤角质细胞HaCaT分为对照组和照射组,照射组给予50 J/m2的UVB照射.照射后培养不同时间,分析miRNA的表达谱.实时荧光定量PCR分析HaCaT细胞和皮肤鳞癌细胞系A431、Tca8113和HSC-1的miR-365的表达.平板克隆形成实验和Transwell小室细胞运动实验分别检测细胞的克隆形成能力和体外运动能力.构建miR-365高表达的HaCaT细胞系HaC aTPre-miR-365-2.裸鼠皮下注射HaCaTpre-miR-365-2观察其成瘤能力.结果 UVB照射后,HaCaT差异表达的miRNAs共30个,其中miR-365最敏感,且照后6h升高最明显,为对照的6.7倍;皮肤鳞癌细胞系A431、Tca8113和HSC-1的miR-365的表达均高于HaCaT细胞(P＜0.05),其中,A431增加最多,为(15.67±1.12)倍,Tca8113最少,为(4.72 ±0.85)倍;抑制皮肤鳞癌细胞系miR-365的表达可以显著抑制其克隆形成能力(=13.68,P＜0.05)和体外细胞迁移能力(t=19.98,P＜0.05),而提高HaCaT的miR-365的表达则可以显著提高其克隆形成能力(t=7.11,P＜0.05)和体外细胞迁移能力(t=22.03,P＜0.05),且能够在裸鼠皮下成功地成瘤.结论 miR-365是一种皮肤鳞状细胞癌的促癌基因.
目的 探討紫外線(UVB)輻射敏感miR-365在皮膚鱗狀細胞癌髮生中的作用.方法 取對數生長期正常人皮膚角質細胞HaCaT分為對照組和照射組,照射組給予50 J/m2的UVB照射.照射後培養不同時間,分析miRNA的錶達譜.實時熒光定量PCR分析HaCaT細胞和皮膚鱗癌細胞繫A431、Tca8113和HSC-1的miR-365的錶達.平闆剋隆形成實驗和Transwell小室細胞運動實驗分彆檢測細胞的剋隆形成能力和體外運動能力.構建miR-365高錶達的HaCaT細胞繫HaC aTPre-miR-365-2.裸鼠皮下註射HaCaTpre-miR-365-2觀察其成瘤能力.結果 UVB照射後,HaCaT差異錶達的miRNAs共30箇,其中miR-365最敏感,且照後6h升高最明顯,為對照的6.7倍;皮膚鱗癌細胞繫A431、Tca8113和HSC-1的miR-365的錶達均高于HaCaT細胞(P＜0.05),其中,A431增加最多,為(15.67±1.12)倍,Tca8113最少,為(4.72 ±0.85)倍;抑製皮膚鱗癌細胞繫miR-365的錶達可以顯著抑製其剋隆形成能力(=13.68,P＜0.05)和體外細胞遷移能力(t=19.98,P＜0.05),而提高HaCaT的miR-365的錶達則可以顯著提高其剋隆形成能力(t=7.11,P＜0.05)和體外細胞遷移能力(t=22.03,P＜0.05),且能夠在裸鼠皮下成功地成瘤.結論 miR-365是一種皮膚鱗狀細胞癌的促癌基因.
목적 탐토자외선(UVB)복사민감miR-365재피부린상세포암발생중적작용.방법 취대수생장기정상인피부각질세포HaCaT분위대조조화조사조,조사조급여50 J/m2적UVB조사.조사후배양불동시간,분석miRNA적표체보.실시형광정량PCR분석HaCaT세포화피부린암세포계A431、Tca8113화HSC-1적miR-365적표체.평판극륭형성실험화Transwell소실세포운동실험분별검측세포적극륭형성능력화체외운동능력.구건miR-365고표체적HaCaT세포계HaC aTPre-miR-365-2.라서피하주사HaCaTpre-miR-365-2관찰기성류능력.결과 UVB조사후,HaCaT차이표체적miRNAs공30개,기중miR-365최민감,차조후6h승고최명현,위대조적6.7배;피부린암세포계A431、Tca8113화HSC-1적miR-365적표체균고우HaCaT세포(P＜0.05),기중,A431증가최다,위(15.67±1.12)배,Tca8113최소,위(4.72 ±0.85)배;억제피부린암세포계miR-365적표체가이현저억제기극륭형성능력(=13.68,P＜0.05)화체외세포천이능력(t=19.98,P＜0.05),이제고HaCaT적miR-365적표체칙가이현저제고기극륭형성능력(t=7.11,P＜0.05)화체외세포천이능력(t=22.03,P＜0.05),차능구재라서피하성공지성류.결론 miR-365시일충피부린상세포암적촉암기인.
Objective To investigate the carcinogeic role of miR-365 in cuntanerous squamous cell carcinoma (cSCC).Methods Normal HaCaT cells were divided into control and irradiation groups,the later was exposed by UVB irradiation (50 J/m2).MicroRNA expression profiles of the two groups were analyzed by microRNA array.The expression variations of miR-365 in HaCaT,A431,Tca8113 and HSC-1 cells were validated by qRT-PCR analysis.The colony-forming and invasion capacities were dectected by colony forming assay and Transwell migration assay in vitro,respectively.HaCaTpre-miR365-2 highly expressing miR-365 was constructed by retroviral vector infection.Tumorigenicity evaluation was carried out by subcutaneously inject of the cells at the right back flank of nude mice.Results There were 30 microRNAs differentially expressed in HaCaT cells after UVB irradiation and miR-365 was one of the most sensitive miRNAs(as high 6.7 times as control).Expression of miR-365 in all the cSCC cell lines A431,Tca8113 and HSC-1 were significantly higher than that in HaCaT cell,in which the maximum was A431 (15.67 ±1.12 times,P ＜ 0.01),and the minimum was TcaS113 (4.72 ± 0.85 times,P ＜ 0.05).Knockdown of miR-365 in cSCC cell lines significantly inhibited the colony forming ability (t =13.68,P ＜ 0.05) and cell migration (t =19.98,P ＜ 0.05) in vitro.HaCaT cells overexpressing miR-365 by transient transfection significantly increased the ability of colony formation (t =7.11,P ＜ 0.05) and cell migration (t =22.03,P ＜0.05) in vitro.In addition,HaCaTpre-miR-365-2 cell line stably expressing miR-365 could successfully establish tumors in nude mice.Conclusions MiR-365 is an oncogene for cutaneous squamous cell carcinoma.
