Survivin反义寡核苷酸选择性诱导肿瘤细胞凋亡研究

We have successively expressed survivin protein in prokaryotic system. The purified recombinant protein was used to create monoclonal antibody by hybridomas. A HeLa cell line which expressed antisense RNA of survivin with the induction of 2mmol/L ZnCl2 has been constructed. Proliferation of HeLa cells was suppressed significantly after the expression of survivin has been inhibited. The inhibition also sensitized HeLa cells to chemothreapeutics and suppressed its cell cycle. It revealed that survivin was an ideal cancertherapy target. It was interesting that inhibiting the expression of telomerase catalytic subunit (TERT) promoted the degradation of survivin protein by ubiquitin pathway. The secondary structure of survivin was predicted by the computer mRNA secondary structure prediction software RNAStructure 3.5. Twenty antisense oligonucleotides (ASON) were designed according to the predicted survivin mRNA secondary structure. ASON No.45 was singled out as the best one by in vitro selection in HeLa S3 cells. It inhibited the proliferation of HeLa S3 cells for more than 50% in 48h. Northern blotting showed that it reduced the mRNA of survivin in HeLa cells. Its reduction of survivin protein correlated well with the concentration of ASON. These results showed that ASON No.45 targeted survivin specifically. It also sensitized HeLa cells to chemotherapeutics harringtonine. ASON No.45 was a promising antisense drug in cancer therapy.And co-inhibiting the expression of TERT and survivin would co-efficiently inhibited the proliferation of cancer cell.

依据Survivin是肿瘤选择性治疗的理想靶标，建立Survivin抑制凋亡活性及其mRNA定量检测技术，计算机辅助设计针对Survivin的反义寡核苷酸（ASON），并进一步评价其体内、外抗肿瘤活性。本课题的实施，对Survivin功能研究，以及以Survivin为靶的抗肿瘤药物设计、研究和有重要的指导意义。