中华医学遗传学杂志
中華醫學遺傳學雜誌
중화의학유전학잡지
CHINESE JOURNAL OF MEDICAL GENETICS
2001年
2期
128-131
,共4页
王柏秋%闫承慧%吴焱%高慧%王琦%金焰%黄承滨%张贵寅%傅松滨%李璞
王柏鞦%閆承慧%吳焱%高慧%王琦%金燄%黃承濱%張貴寅%傅鬆濱%李璞
왕백추%염승혜%오염%고혜%왕기%금염%황승빈%장귀인%부송빈%리박
p16基因%p21基因%p53基因%转染%基因治疗%肺腺癌
p16基因%p21基因%p53基因%轉染%基因治療%肺腺癌
p16기인%p21기인%p53기인%전염%기인치료%폐선암
目的探讨肿瘤抑制基因对肺腺癌细胞生长的抑制作用。方法利用FuGene转染方式分别将p21和p16基因的表达质粒转入一对肺腺癌细胞系Anip973和AGZY83-a中，同时用含野生型p53基因的腺病毒感染p16基因转染前后的这一对细胞系。对P16和P21蛋白过表达的细胞系进行了细胞生长曲线、克隆形成率、原位末端标记分析和流式细胞仪分析。结果 p16基因的过表达只能使细胞系的G1期细胞比例提高，但细胞生长曲线、克隆形成率均未出现改变，未检测到凋亡信号。P21蛋白过表达的一对细胞系细胞生长曲线斜率降低，克隆形成能力下降，并出现明显的G1期阻滞，但未检测到凋亡信号。p53基因感染AGZY83-a、Anip973及经过p16基因转染的细胞AGZY83-ap16和Anip973p16后呈现时间依赖性表达，细胞生长曲线和四唑盐比色法分析提示，野生型p53基因的大量表达明显抑制以上4种细胞的生长，Anip973和Anip973p16的生长抑制率高于AGZY83-a和AGZY83-ap16；Anip973p16和AGZY83-ap16的生长抑制率高于Anip973和AGZY83-a。这4种细胞在感染p53后出现典型的凋亡信号。结论 p16基因的过表达并不能抑制细胞系的生长，而p21基因的过表达通过G1期阻滞抑制这1对肺腺癌细胞的生长；野生型p53基因在AGZY83-a和Anip973中高效表达可产生明显的细胞生长抑制效应；野生型p53基因对肺腺癌高转移细胞系Anip973抑制作用更为明显。
目的探討腫瘤抑製基因對肺腺癌細胞生長的抑製作用。方法利用FuGene轉染方式分彆將p21和p16基因的錶達質粒轉入一對肺腺癌細胞繫Anip973和AGZY83-a中，同時用含野生型p53基因的腺病毒感染p16基因轉染前後的這一對細胞繫。對P16和P21蛋白過錶達的細胞繫進行瞭細胞生長麯線、剋隆形成率、原位末耑標記分析和流式細胞儀分析。結果 p16基因的過錶達隻能使細胞繫的G1期細胞比例提高，但細胞生長麯線、剋隆形成率均未齣現改變，未檢測到凋亡信號。P21蛋白過錶達的一對細胞繫細胞生長麯線斜率降低，剋隆形成能力下降，併齣現明顯的G1期阻滯，但未檢測到凋亡信號。p53基因感染AGZY83-a、Anip973及經過p16基因轉染的細胞AGZY83-ap16和Anip973p16後呈現時間依賴性錶達，細胞生長麯線和四唑鹽比色法分析提示，野生型p53基因的大量錶達明顯抑製以上4種細胞的生長，Anip973和Anip973p16的生長抑製率高于AGZY83-a和AGZY83-ap16；Anip973p16和AGZY83-ap16的生長抑製率高于Anip973和AGZY83-a。這4種細胞在感染p53後齣現典型的凋亡信號。結論 p16基因的過錶達併不能抑製細胞繫的生長，而p21基因的過錶達通過G1期阻滯抑製這1對肺腺癌細胞的生長；野生型p53基因在AGZY83-a和Anip973中高效錶達可產生明顯的細胞生長抑製效應；野生型p53基因對肺腺癌高轉移細胞繫Anip973抑製作用更為明顯。
목적탐토종류억제기인대폐선암세포생장적억제작용。방법이용FuGene전염방식분별장p21화p16기인적표체질립전입일대폐선암세포계Anip973화AGZY83-a중，동시용함야생형p53기인적선병독감염p16기인전염전후적저일대세포계。대P16화P21단백과표체적세포계진행료세포생장곡선、극륭형성솔、원위말단표기분석화류식세포의분석。결과 p16기인적과표체지능사세포계적G1기세포비례제고，단세포생장곡선、극륭형성솔균미출현개변，미검측도조망신호。P21단백과표체적일대세포계세포생장곡선사솔강저，극륭형성능력하강，병출현명현적G1기조체，단미검측도조망신호。p53기인감염AGZY83-a、Anip973급경과p16기인전염적세포AGZY83-ap16화Anip973p16후정현시간의뢰성표체，세포생장곡선화사서염비색법분석제시，야생형p53기인적대량표체명현억제이상4충세포적생장，Anip973화Anip973p16적생장억제솔고우AGZY83-a화AGZY83-ap16；Anip973p16화AGZY83-ap16적생장억제솔고우Anip973화AGZY83-a。저4충세포재감염p53후출현전형적조망신호。결론 p16기인적과표체병불능억제세포계적생장，이p21기인적과표체통과G1기조체억제저1대폐선암세포적생장；야생형p53기인재AGZY83-a화Anip973중고효표체가산생명현적세포생장억제효응；야생형p53기인대폐선암고전이세포계Anip973억제작용경위명현。
Objective In order to investigate the suppression effect of tumor suppressor genes in lung adenocarcinoma. Methods p16 and p21 expression vectors were transfected into a pair of lung adenocarcinoma cell lines with different metastasis potentials:Anip973(high metastasis potential)and AGZY83-a (low metastasis potential). In the mean time, AGZY83-a, Anip973, AGZY83-ap16 and Anip973p16 were infected with recombinant adenovirus encoding wild-type p53 gene. The suppression effects of these genes were evaluated by cell growth curve, MTT,cloning efficiency assay, flow cytometric analysis and TUNEL technique. Results Overexpression of p16 gene in Anip973 and AGZY83-a could only lengthen the G1 phase while increased expression of p21 in both of the cell lines was associated with significant lengthening of G1 phase, decreased proliferation potential and decreased cloning efficiency. High efficient expression of wild-type p53 gene in AGZY83-a, Anip973, Anip973p16 and AGZY83-ap16 inhibited the growth of these four kinds of lung cancer cells and killed the cells in the end. Apoptosis was detected in all the four kinds of cells. The suppression effect of p53 gene was higher in Anip973 and Anip973p16 than in AGZY83-a and AGZY83-ap16 while co-expression of p53 and p16 in this pair of cell lines inhibited the cells more efficiently as compared with the expression of p53 gene. Conclusion Increased expression of p21 gene suppressed the lung adenocarcinoma cells by G1 arrest and the co-transfection of tumor suppressor genes p16 and p53 into the lung adenocarcinoma cell line proved more effective in lung cancer gene therapy.
