中华医学杂志(英文版)
中華醫學雜誌(英文版)
중화의학잡지(영문판)
CHINESE MEDICAL JOURNAL
2001年
5期
497-501
,共5页
白景香%祝学光%郑新程%伍贻经
白景香%祝學光%鄭新程%伍貽經
백경향%축학광%정신정%오이경
p16基因逆转录病毒载体乳腺癌细胞细胞周期阻滞
p16基因逆轉錄病毒載體乳腺癌細胞細胞週期阻滯
p16기인역전록병독재체유선암세포세포주기조체
目的研究p16基因对肿瘤细胞生长抑制及细胞周期阻滞作用。方法将p16 cDNA插入逆转录病毒载体pLXSN构建成p16基因逆转录病毒重组体pLp16SN。利用基因转染方法，将pLp16SN及pLXSN导人逆转录病毒包装细胞系PA317细胞，获得逆转录病毒。用逆转录病毒感染Bcap-37乳腺癌细胞，经G418筛选获阳性克隆。利用Northern和Western blotting方法检测p16基因的表达。检测转基因细胞的生长速度，细胞周期及裸鼠成瘤等细胞生物学行为的改变。结果 Northem及Western blotting显示转染p16基因的Bcap-37细胞p16基因mRNA及蛋白质表达明显增高。此细胞较未转染基因的Bcap-37细胞和转染空载体的Bcap-37细胞生长速度慢，G1期细胞比率增高，裸鼠接种成瘤体积小。结论 p16基因高表达能够抑制乳腺癌细胞Bcap-37的生长，并阻滞细胞从G1期进入S期。
目的研究p16基因對腫瘤細胞生長抑製及細胞週期阻滯作用。方法將p16 cDNA插入逆轉錄病毒載體pLXSN構建成p16基因逆轉錄病毒重組體pLp16SN。利用基因轉染方法，將pLp16SN及pLXSN導人逆轉錄病毒包裝細胞繫PA317細胞，穫得逆轉錄病毒。用逆轉錄病毒感染Bcap-37乳腺癌細胞，經G418篩選穫暘性剋隆。利用Northern和Western blotting方法檢測p16基因的錶達。檢測轉基因細胞的生長速度，細胞週期及裸鼠成瘤等細胞生物學行為的改變。結果 Northem及Western blotting顯示轉染p16基因的Bcap-37細胞p16基因mRNA及蛋白質錶達明顯增高。此細胞較未轉染基因的Bcap-37細胞和轉染空載體的Bcap-37細胞生長速度慢，G1期細胞比率增高，裸鼠接種成瘤體積小。結論 p16基因高錶達能夠抑製乳腺癌細胞Bcap-37的生長，併阻滯細胞從G1期進入S期。
목적연구p16기인대종류세포생장억제급세포주기조체작용。방법장p16 cDNA삽입역전록병독재체pLXSN구건성p16기인역전록병독중조체pLp16SN。이용기인전염방법，장pLp16SN급pLXSN도인역전록병독포장세포계PA317세포，획득역전록병독。용역전록병독감염Bcap-37유선암세포，경G418사선획양성극륭。이용Northern화Western blotting방법검측p16기인적표체。검측전기인세포적생장속도，세포주기급라서성류등세포생물학행위적개변。결과 Northem급Western blotting현시전염p16기인적Bcap-37세포p16기인mRNA급단백질표체명현증고。차세포교미전염기인적Bcap-37세포화전염공재체적Bcap-37세포생장속도만，G1기세포비솔증고，라서접충성류체적소。결론 p16기인고표체능구억제유선암세포Bcap-37적생장，병조체세포종G1기진입S기。
Objective To study the effects of the p16 gene on tumor cell growth inhibition and cell cycle arrest. Methods The recombinant retroviral vector pLp16SN was constructed by cloning the human p16 cDNA into the retroviral vector pLXSN. Retroviruses with or without the p16 gene were obtained by transfecting pLp16SN and pLXSN vectors into PA317 cells. Bcap-37 human breast cancer cells were infected with these retroviruses followed by selection with G418. The expression of p16 was detected by Northern and Westem blots. Cell biological characteristics, including cell growth rate, cell cycle and tumorigenesis in nude mice were assessed.Results Both mRNA and protein expression of p16 in Bcap-37 cells transfected with retroviral vector containing the p16 gene were much higher than that in Bcap-37 cells transfected with empty vector or parental Bcap-37 cells. Cell overexpressing the p16 gene exhibited a slower rate of growth, a higher percentage of cells in the G1 phase, and smaller tumors in nude mice, compared with parental Bcap-37 cells and cells transfected with empty vector.Conclusion Overexpression of the p16 gene could suppress the growth of Bcap-37 breast cancer cells by arresting the cell cvcle at the G1 to S-phase.
