Gu Xia, Lin Han-Liang, Shao Jian-Yong, Zhang Meng, Zhu You-Kai, Liang Hui-Zhen, Ma Yi-Hui
Department of Pathology, Basic Medical College, Sun Yat-sen University, Guangzhou 510089, China.
Zhonghua Bing Li Xue Za Zhi. 2005 Nov;34(11):737-41.
To study the effect of transfecting survivin antisense mRNA on growth and chemotherapy sensitivity of lymphoma cells.
Eukaryotic expression plasmid pcDNA3. 1-antisense (As) survivin was constructed and transfected into Jurkat T lymphoblastic lymphoma cell lines with high expression survivin mRNA by use of lipofectmine gene transfer technique. Expression of survivin mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemical and Western blot. The effect of transfecting survivin antisense mRNA on the growth of Jurkat cell lines was monitored by population doubling time (PDT) and Apoptotic indexes (AI). The morphologic features were observed in transfected cells by light and electric microscopes. MTT assay was used to analyze the response of transfected cells to CTX and MTX.
Compared with the control cells, the expression of survivin mRNA and protein were reduced after transfected pcDNA3. 1-Assurvivin 48 h, 5 w and 6 w, PDT (52 h) was prolonged. Apoptotic indexes were higher in transfected antisense survivin mRNA cells [20.2% (48 h)], 6.2% (5 w) and 6.8% (6 w) than control ones [2.1%, 1.3% (48 h)] and [1.3% (5 w) and 1.0% (6 w)]. The cells grow slowly and the dead cells increase and some swelling and apoptotic cells were observed in transfected pcDNA3. 1-Assurvivin groups by invert, light and electric microscopes. The Jurkat cell line of transfected pcDNA3. 1-Assurvivin had higher sensitivity to CTX and MTX. The rate of inhibition was higher in transfected group. There is a significant difference between the transfected group and untransfected one, P < 0.05.
The result indicated that survivin gene was very important for growth of Jurkat cells. To inhibit the expression of survivin will be significant in therapy of T lymphoblastic lymphoma. Survivin gene might be a target of therapy.
研究转染survivin反义mRNA对淋巴瘤细胞生长及化疗敏感性的影响。
构建真核表达质粒pcDNA3.1-反义（As）survivin，采用脂质体基因转染技术将其转染入survivin mRNA高表达的Jurkat T淋巴细胞淋巴瘤细胞系。通过逆转录-聚合酶链反应（RT-PCR）、免疫组织化学及蛋白质印迹法检测survivin mRNA和蛋白的表达。通过群体倍增时间（PDT）和凋亡指数（AI）监测转染survivin反义mRNA对Jurkat细胞系生长的影响。利用光学显微镜和电子显微镜观察转染细胞的形态学特征。采用MTT法分析转染细胞对CTX和MTX的反应。
与对照细胞相比，转染pcDNA3.1-As survivin 48小时、5周和6周后，survivin mRNA和蛋白的表达降低，PDT（52小时）延长。转染反义survivin mRNA的细胞凋亡指数[20.2%（48小时）]、6.2%（5周）和6.8%（6周）高于对照细胞[2.1%、1.3%（48小时）]和[1.3%（5周）和1.0%（6周）]。通过倒置显微镜、光学显微镜和电子显微镜观察，转染pcDNA3.1-As survivin组细胞生长缓慢，死亡细胞增多，可见一些肿胀和凋亡细胞。转染pcDNA3.1-As survivin的Jurkat细胞系对CTX和MTX具有更高的敏感性。转染组的抑制率更高。转染组与未转染组之间存在显著差异，P<0.05。
结果表明survivin基因对Jurkat细胞的生长非常重要。抑制survivin的表达对T淋巴细胞淋巴瘤的治疗具有重要意义。Survivin基因可能是治疗靶点。
