抑瘤基因NGX6对人结肠癌细胞HT-29生长的影响

图1 重组体质粒双酶切鉴定结果. M: DL2000Marker; 1-5为载体重组体酶切后产物均为阳性克隆.

图2 pcDNA3. 1(+)/NGX6转染HT-29细胞抗性克隆的RT-PCR鉴定结果. M: DL 2 000 marker; 空白为未转染的HT-29克隆; +2~+12为转染pcDNA3.1(+)/NGX6重组体的克隆, 其中+2, +10为阳性克隆.

图3 pcDNA3. 1(+)转染HT-29细胞抗性克隆的RT-PCR鉴定结果. M: DL 2000 marker; 空白: 为未转染HT-29组; -3~-6为pcDNA3.1(+)转染组均阳性; -4, -6克隆pcDNA3.1(+)表达量高.

图4 pcDNA3. 1(+)/NGX6和pcDNA3.1(+)转染的HT-29抗性克隆分别以NGX6, GAPDH为探针的点杂交鉴定结果.

图5 NGX6基因转染前后的HT-29细胞生长曲线.

图6 NGX6转染对结肠癌细胞HT-29倍增时间的影响.

图7 NGX6转染前后HT-29细胞的软琼脂集落形成. A: HT-29组(×100); B: HT-29组(×400)C: pcDNA3.1(+)组(×100); D: pcDNA3.1(+)组(×400)E: pcDNA3.1(+)/NGX6组(×100); F: pcDNA3.1(+)/NGX6组(×400)G: HT-29和pcDNA3.1(+)/HT-29组克隆数多, 体积大; pcDNA3.1(+)/NGX6/HT-29组克隆数明显减少, 体积小.

图8 NGX6转染前后裸鼠移植瘤体积变化.

图9 pcDNA3. 1(+)/HT-29和pcDNA3.1(+)/NGX6/HT-29组移植瘤HE(×200); A pcDNA3.1(+)/HT-29组移植瘤可见瘤体包膜断裂、不完整, 箭头所示处血管瘤栓形成; B pcDNA3.1(+)/NGX6/HT-29组移植瘤内血管壁完整, 未见瘤细胞侵犯.