Guan Hai-tao, Xue Xing-huan, Wang Xi-jing, Li Ang, Qin Zhao-yin
Department of Oncologic Surgery, Second Affiliated Hospital of Xi'an Jiaotong University Medical College, Xi'an 710004, China.
Zhonghua Zhong Liu Za Zhi. 2006 May;28(5):326-30.
Blocking the expression of survivin with RNA interference techniques, the effects of suppressing proliferation and inducing apoptosis of breast cancer MCF-7 cells were investigated.
A siRNA eukaryotic expression vector against survivin was constructed and transfected into breast cancer MCF-7 cells with lipofectamine 2000. The changes of survivin expression were detected by semi-quantitive RT-PCR and immunohistochemistry. The effect of suppressing proliferation of MCF-7 cell was detected by MTT assay. The effect of inducing MCF-7 cell apoptosis was detected by TUNEL assay.
The sequence-specific siRNA can efficiently block the expression of survivin both at mRNA and protein levels. The expression inhibition rate was 64.9% at mRNA level detected by semi-quantitive RT-PCR and 79.7% at protein level detected by immunohistochemistry. Blocking the expression of survivin can suppress proliferation of MCF-7 cells significantly. At 24 and 48 h after the cells were reseeded, the proliferation inhibition rates were 31.6% and 33.0%, respectively. At 24 h after transfection, apoptosis was induced in 12.9% of the cells as detected by TUNEL assay.
Blocking the expression of survivin with RNA interference technology can significantly suppress proliferation of MCF-7 cells and induce apoptosis to a certain degree. RNAi targeted to survivin has a potential value in gene therapy of breast cancer.
运用RNA干扰技术阻断生存素的表达,研究其对乳腺癌MCF-7细胞增殖抑制及诱导凋亡的作用。
构建针对生存素的siRNA真核表达载体,并用脂质体2000转染乳腺癌MCF-7细胞。采用半定量RT-PCR和免疫组织化学检测生存素表达的变化。用MTT法检测对MCF-7细胞增殖的抑制作用。用TUNEL法检测诱导MCF-7细胞凋亡的作用。
序列特异性siRNA能在mRNA和蛋白水平有效阻断生存素的表达。半定量RT-PCR检测mRNA水平的表达抑制率为64.9%,免疫组织化学检测蛋白水平的表达抑制率为79.7%。阻断生存素的表达能显著抑制MCF-7细胞的增殖。细胞重新接种后24小时和48小时,增殖抑制率分别为31.6%和33.0%。转染后24小时,TUNEL法检测发现12.9%的细胞发生凋亡。
运用RNA干扰技术阻断生存素的表达能显著抑制MCF-7细胞的增殖并在一定程度上诱导凋亡。针对生存素的RNA干扰在乳腺癌基因治疗中具有潜在价值。