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目的通过shRNA沉默吲哚胺2,3-双加氧酶(indoleamine2,3-dioxygenase,IDO)基因表达,研究IDO表达在体外对NK细胞杀伤能力的作用。方法 shRNA沉默IDO基因表达质粒和空白质粒分别稳定转染至人卵巢癌细胞SKOV-3,应用Western blot检测IDO在SKOV-3、SKOV-3/Mock和SKOV-3/shIDO细胞中的表达情况,用MTT试剂盒检测3组肿瘤细胞体外生长速度和对NK细胞杀伤作用的敏感性。结果 IDO蛋白在SKOV-3/shIDO细胞中表达被抑制,在SKOV-3和SKOV-3/Mock细胞中有表达。3组肿瘤细胞体外生长曲线比较差异无统计学意义(P>0.05)。SKOV-3/shIDO细胞存活的百分比明显低于其他2组对照(SKOV-3和SKOV-3/Mock)细胞,差异有统计学意义(P<0.05)。结论本研究应用shRNA沉默IDO基因表达质粒稳定转染卵巢癌细胞SKOV-3,获得IDO无表达卵巢癌细胞SKOV-3/shIDO,结果显示抑制IDO表达对卵巢癌细胞体外生长速度无影响,但可增强卵巢癌细胞SKOV-3对NK细胞杀伤作用的敏感性。因此,IDO可以作为卵巢癌基因治疗的潜在新靶点,shRNA沉默IDO基因表达可以作为卵巢癌治疗的新方法。
Objective To investigate the effect of IDO expression on cytotoxicity of NK cells in vitro by shRNA expression of indoleamine 2,3-dioxygenase (IDO) gene. Methods shRNA-IDO gene expression plasmid and blank plasmid were stably transfected into human ovarian cancer cell line SKOV-3. Western blot was used to detect the expression of IDO in SKOV-3, SKOV-3 / Mock and SKOV-3 / The MTT kit was used to detect the growth rate of tumor cells in vitro and the sensitivity to NK cell killing. Results IDO protein was inhibited in SKOV-3 / shIDO cells and expressed in SKOV-3 and SKOV-3 / Mock cells. The growth curve of tumor cells in three groups showed no significant difference (P> 0.05). The percentage of SKOV-3 / shIDO cells was significantly lower than that of other two groups (SKOV-3 and SKOV-3 / Mock). The difference was statistically significant (P <0.05). Conclusions In this study, we successfully transfected ovarian cancer cell line SKOV-3 with shRNA silencing IDO gene expression plasmid to obtain IDO non-expressing ovarian cancer cell line SKOV-3 / shIDO. The results showed that inhibition of IDO expression had no effect on the growth rate of ovarian cancer cell in vitro, Enhance the sensitivity of SKOV-3 to kill NK cells in ovarian cancer cells. Therefore, IDO can be used as a potential new target for ovarian cancer gene therapy. ShRNA silencing of IDO gene expression can be used as a new treatment for ovarian cancer.