中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2009年
6期
423-426
,共4页
徐文贵%杨昆%房娜%戴东%于津浦%魏枫%宋秀宇%朱研佳%王健%门晓媛%张莹
徐文貴%楊昆%房娜%戴東%于津浦%魏楓%宋秀宇%硃研佳%王健%門曉媛%張瑩
서문귀%양곤%방나%대동%우진포%위풍%송수우%주연가%왕건%문효원%장형
乳腺肿瘤%肿瘤移植%自杀基因%T47D-tk
乳腺腫瘤%腫瘤移植%自殺基因%T47D-tk
유선종류%종류이식%자살기인%T47D-tk
Breast neoplasms%Noplasm transplantation%Suicide gene%T47D-tk
目的 进一步对已经获得的稳定表达自杀基因HSV1-tk的乳腺癌细胞株T47D-tk进行鉴定,观察其生长特性,建立移植瘤动物模型.方法 提取细胞基因组、以HSV1-tk P1和HSV1-tk P2为引物,进行PCR反应,琼脂糖凝胶电泳基因鉴定.描绘生长曲线、镜下观察细胞形态.BALB/c nu/nu裸鼠前肢皮下接种T47D和T47D-tk细胞,并观察二者的成瘤特性.结果 T47D-tk细胞基因组DNA进行PCR检测可见到大小约1131 bp的片段,而正常的T47D细胞基因组则无此阳性条带出现.两组平均生长细胞数,第1天T47D:(5.58±1.04)×103,T47D-tk:(6.08±1.61)×103;第3天T47D:(10.00±1.30)×103;T47D-tk:(10.25±0.90)×103;第7天T47D:(19.25±0.66)×103,T47D-tk:(19.00±1.80)×103,两组间差异均无统计学意义(均P>0.05),形态观察亦未见明显差别.接种后两组成瘤时间[T47D:(9.67±0.33)d;T47D-tk:(9.83±0.48)d]和肿瘤大小差异均无统计学意义(均P>0.05).结论 T47D-tk细胞基因组中已整合入HSV1-tk基因,与T47D细胞相比生长特性无变化,成功建立了T47D和T47D-tk移植瘤动物模型,为进一步基因显像与基因治疗奠定了基础.
目的 進一步對已經穫得的穩定錶達自殺基因HSV1-tk的乳腺癌細胞株T47D-tk進行鑒定,觀察其生長特性,建立移植瘤動物模型.方法 提取細胞基因組、以HSV1-tk P1和HSV1-tk P2為引物,進行PCR反應,瓊脂糖凝膠電泳基因鑒定.描繪生長麯線、鏡下觀察細胞形態.BALB/c nu/nu裸鼠前肢皮下接種T47D和T47D-tk細胞,併觀察二者的成瘤特性.結果 T47D-tk細胞基因組DNA進行PCR檢測可見到大小約1131 bp的片段,而正常的T47D細胞基因組則無此暘性條帶齣現.兩組平均生長細胞數,第1天T47D:(5.58±1.04)×103,T47D-tk:(6.08±1.61)×103;第3天T47D:(10.00±1.30)×103;T47D-tk:(10.25±0.90)×103;第7天T47D:(19.25±0.66)×103,T47D-tk:(19.00±1.80)×103,兩組間差異均無統計學意義(均P>0.05),形態觀察亦未見明顯差彆.接種後兩組成瘤時間[T47D:(9.67±0.33)d;T47D-tk:(9.83±0.48)d]和腫瘤大小差異均無統計學意義(均P>0.05).結論 T47D-tk細胞基因組中已整閤入HSV1-tk基因,與T47D細胞相比生長特性無變化,成功建立瞭T47D和T47D-tk移植瘤動物模型,為進一步基因顯像與基因治療奠定瞭基礎.
목적 진일보대이경획득적은정표체자살기인HSV1-tk적유선암세포주T47D-tk진행감정,관찰기생장특성,건립이식류동물모형.방법 제취세포기인조、이HSV1-tk P1화HSV1-tk P2위인물,진행PCR반응,경지당응효전영기인감정.묘회생장곡선、경하관찰세포형태.BALB/c nu/nu라서전지피하접충T47D화T47D-tk세포,병관찰이자적성류특성.결과 T47D-tk세포기인조DNA진행PCR검측가견도대소약1131 bp적편단,이정상적T47D세포기인조칙무차양성조대출현.량조평균생장세포수,제1천T47D:(5.58±1.04)×103,T47D-tk:(6.08±1.61)×103;제3천T47D:(10.00±1.30)×103;T47D-tk:(10.25±0.90)×103;제7천T47D:(19.25±0.66)×103,T47D-tk:(19.00±1.80)×103,량조간차이균무통계학의의(균P>0.05),형태관찰역미견명현차별.접충후량조성류시간[T47D:(9.67±0.33)d;T47D-tk:(9.83±0.48)d]화종류대소차이균무통계학의의(균P>0.05).결론 T47D-tk세포기인조중이정합입HSV1-tk기인,여T47D세포상비생장특성무변화,성공건립료T47D화T47D-tk이식류동물모형,위진일보기인현상여기인치료전정료기출.
Objective To further identify the breast adenocarcinoma cell line T47D-tk stably expressing the suicide gene HSV1-tk,observe its growing characteristics,and establish an animal model of implanted breast adenocarcinoma. Methods Retroviral vector of HSV1-tk gene and breast carcinoma cell line T47D-tk stably expressing the HSV1-tk gene were established. Breast carcinoma cells of the lines T47D and T47D-tk were cultured,and observed by inverted microscope. Growth curve was drawn. Genomic DNA of T47D-tk cells was extracted,and amplified by PCR with HSV1-tk and HSV1-tk P2 as primers. Agarose gel electrophoresis was used to identify the HSV1tk gene. Suspensions of T47D and T47D-tk cells were inoculated subcutaneously at bilateral roots of foreleg of female BALB/c nude mice respectively. The growth of tumor was observed every day. Results PCR showed 1131 bp fragment in the T47 D-tk genome,but not in the T47D genome. The numbers of growing T47D cells on days 3,and 7 were ( 10.00 ± 1,30) × 103 and ( 19.25±0.66) × 103 respectively,not significantly different from those of the T47D-tk cells [(10. 25 ±0.90) × 103 and ( 19.00 ± 1.80) × 103 respectively,both P > 0. 05]. The time needed for tumor formation after the inoculation of T47D cells was (9.67 ± 0.33 )d,not significantly different from that of the T47D-tk cells [ (9.83±0.48) d,P >0.05 ]. The tumor size 19 days after inoculation of the T47D cells was (72. 17±25.88) mm3,not significantly different from that of the T47D-tk cells [ (70.66±22.16) mm3,P >0.05 ]. Conclusion The T47D-tk cells have integrated the HSV1-tk gene without changing its growing characteristics. An animal model of implanted breast cancer has been successfully established. The T47D and T47D-tk subcutaneous xenografts offer the foundation for further study of gene imaging and gene therapy.
