中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2008年
6期
395-399
,共5页
马儒林%庞雅琴%李文学%肖勇梅%魏青%李道传%赖延东%林育纯%王庆%杨萍%陈丽萍%唐石伏%林忠宁%陈雯
馬儒林%龐雅琴%李文學%肖勇梅%魏青%李道傳%賴延東%林育純%王慶%楊萍%陳麗萍%唐石伏%林忠寧%陳雯
마유림%방아금%리문학%초용매%위청%리도전%뢰연동%림육순%왕경%양평%진려평%당석복%림충저%진문
细胞转化,肿瘤%上皮细胞%基因,ras%基因,myc
細胞轉化,腫瘤%上皮細胞%基因,ras%基因,myc
세포전화,종류%상피세포%기인,ras%기인,myc
Cell transformation,neoplastic%Epithelial cells%Genes,ras%Genes,myc
目的 构建癌基因高表达人支气管上皮细胞模型并应用于致癌物诱导细胞恶性转化活性的检测.方法 在永生化人支气管上皮细胞株16HBE中依次导入人端粒酶催化亚基(hTERT)、癌基因H-RasV12或c-Myc或空白载体,构建细胞株16HBETR、16HBETM和16HBETV.检测所构建细胞株的生物学特性如细胞形态、细胞生长、染色体畸变等指标.用已知致癌物硫酸镍(NiSO4)和二氢二醇环氧苯并(a)芘(BPDE)多次染毒,利用软琼脂试验及荷瘤试验检测化学物诱导细胞转化的活性.结果 经过端粒酶活性测定以及蛋白印迹验证上述转基因16HBE细胞株均构建成功.癌基因H-Ras和c-Myc的表达分别使细胞的增殖加速30.3%和10.4%,但是细胞染色体的畸变率没有发生改变.癌基因高表达细胞株只在软琼脂中形成少数背景克隆,但不能在裸鼠皮下形成肿瘤.BPDE诱导16HBETR、16HBETM细胞发生恶性转化的间期分别为5周、11周,比对照组细胞16HBETV的20周分别提前了15周和9周;而硫酸镍诱导16HBETR、16HBETM细胞转化的间期分别为11周、14周.比对照组细胞的32周分别提前了21周和18周.结论 癌基因高表达的细胞转化模型可大大缩短试验间期,有望应用于环境致癌物的筛查及化学致癌机制的研究.
目的 構建癌基因高錶達人支氣管上皮細胞模型併應用于緻癌物誘導細胞噁性轉化活性的檢測.方法 在永生化人支氣管上皮細胞株16HBE中依次導入人耑粒酶催化亞基(hTERT)、癌基因H-RasV12或c-Myc或空白載體,構建細胞株16HBETR、16HBETM和16HBETV.檢測所構建細胞株的生物學特性如細胞形態、細胞生長、染色體畸變等指標.用已知緻癌物硫痠鎳(NiSO4)和二氫二醇環氧苯併(a)芘(BPDE)多次染毒,利用軟瓊脂試驗及荷瘤試驗檢測化學物誘導細胞轉化的活性.結果 經過耑粒酶活性測定以及蛋白印跡驗證上述轉基因16HBE細胞株均構建成功.癌基因H-Ras和c-Myc的錶達分彆使細胞的增殖加速30.3%和10.4%,但是細胞染色體的畸變率沒有髮生改變.癌基因高錶達細胞株隻在軟瓊脂中形成少數揹景剋隆,但不能在裸鼠皮下形成腫瘤.BPDE誘導16HBETR、16HBETM細胞髮生噁性轉化的間期分彆為5週、11週,比對照組細胞16HBETV的20週分彆提前瞭15週和9週;而硫痠鎳誘導16HBETR、16HBETM細胞轉化的間期分彆為11週、14週.比對照組細胞的32週分彆提前瞭21週和18週.結論 癌基因高錶達的細胞轉化模型可大大縮短試驗間期,有望應用于環境緻癌物的篩查及化學緻癌機製的研究.
목적 구건암기인고표체인지기관상피세포모형병응용우치암물유도세포악성전화활성적검측.방법 재영생화인지기관상피세포주16HBE중의차도입인단립매최화아기(hTERT)、암기인H-RasV12혹c-Myc혹공백재체,구건세포주16HBETR、16HBETM화16HBETV.검측소구건세포주적생물학특성여세포형태、세포생장、염색체기변등지표.용이지치암물류산얼(NiSO4)화이경이순배양분병(a)비(BPDE)다차염독,이용연경지시험급하류시험검측화학물유도세포전화적활성.결과 경과단립매활성측정이급단백인적험증상술전기인16HBE세포주균구건성공.암기인H-Ras화c-Myc적표체분별사세포적증식가속30.3%화10.4%,단시세포염색체적기변솔몰유발생개변.암기인고표체세포주지재연경지중형성소수배경극륭,단불능재라서피하형성종류.BPDE유도16HBETR、16HBETM세포발생악성전화적간기분별위5주、11주,비대조조세포16HBETV적20주분별제전료15주화9주;이류산얼유도16HBETR、16HBETM세포전화적간기분별위11주、14주.비대조조세포적32주분별제전료21주화18주.결론 암기인고표체적세포전화모형가대대축단시험간기,유망응용우배경치암물적사사급화학치암궤제적연구.
Objective To establish human bronchial epithelial cell lines over expressing oncogene and to investigate its application in detection of carcinogen—induced cell transformation.Methods Mediated by retrovirus infection,human telomerase catalytic subunit.hTERT was introduced into immortal human bronchial epithelial ceNs(16HBE)and followed by introducfion of the oncogenic allele H—Ras(V12),or c. Myc or empty vector,creating cell lines 16HBETR,16HBETM and 16HBETV.respectively.Biological characteristics of tllese cell lines including morphology.proliferation,and chromosomal aberration were examined to access whether they were transformed.Soft agar experiment and nude mice subcutaneous iniection were performed using pre-transformed 1 6HBE ceUs induced by known carcinogens.nickel sulfate (NiS04)and 7,8,-dihydrodiol-9,10-epoxide benzo[a]pyrene(BPDE).Results witll detection of telomerase activity and Western blotting,the expression of target proteins Was verifed.Thus,the transgenic 16HBE cell lines were successfully established.Cells expressing oncogefie H—Ras or c.Myc grew 30.3%or 10.4%faster than control cells.However.these cells falied to form colonies in soft agar or form tumor in nude mice.16HBETR,16HBETM cells obtained transformed phenotype at 5 wks.11 wks,respectively after treatment with BPDE,which are 15 wks and 9 wks earlier than control cells 16HBETV(20 wks). Meanwhile,16HBETR。16HBETM cells obtained~ansformed phenotype at ll wks,14 wks.respectively after treatment with nickel sulfate,which are 21 wks and 18 wks earlier than control eeUs(32 wks). Condusion With t11e advantage of shorter latency.transgenic human eell transformation models could be used in potent carcinogen screening and applied to chemical—carcinogensis mechanism study.
