Cyclin D1反义寡核苷酸对胃癌细胞化疗敏感性的影响

Effects of Cyclin D1 Antisense Oligodeoxyneucleotides on Chemosensitivity of Gastric Carcinoma Cells

目的研究Cyclin D1反义寡核苷酸(ASODN)对胃癌细胞SGC-7901和HS-746T化疗敏感性的影响,并探讨其内在机制。方法在给予Cyclin D1ASODN和Cyclin D1反义寡核苷酸转染细胞24h后,分别给予梯度浓度的5-氟尿嘧啶(5-FU)、氨甲喋呤(MTX)、顺铂(CDDP),观察各组的量效反应,计算IC50。Cyclin D1ASODN转染细胞后24h、48h时应用RT-PCR分别检测各组细胞中胸苷酸合酶(TS)、胸腺嘧啶磷酸化酶(TP)、二氢叶酸还原酶(DHFR)的表达情况。结果Cyclin D1ASODN转染增加了两种胃癌细胞对5-FU、MTX、CDDP的敏感性,ASODN+化疗组对各化疗药物的IC50与对照组相比均有显著下降。RT-PCR显示Cyclin D1ASODN转染后24h时Cyclin D1、TS、DHFR mRNA表达较对照组下降,而TP mRNA表达较对照组升高,在48h时各种酶mRNA表达的改变更加明显。结论Cyclin D1ASODN能提高胃癌细胞对多种化疗药物的敏感性,可能是由于影响了化疗药物代谢相关酶表达的改变所致。

Objectives To examine the effect of Cyclin D1 antisense oligodeoxyneucleotides （ ASODN ） on chemosensitivity of gastric carcinoma cell lines SGC-7901 and HS-746T. Methods 5-FU, MTX, CDDP in different concentrations were given to every group after transfected with Cyclin D1 SOND or ASODN for 24 h ,the do^-effects response was observed and 1C50 was calculated in every group. The mRNA expressions of Cyclin D1, Thymidylate Synthase （TS） , Thymidine Phosphorylase （TP） , Dihydrofolate Reductase （DHFR） were detected by reverse transcriptionPCR （RT-PCR） in every group at 24 h and 48 h after transfection. Results Cyclin D1 ASODN could increase the sensitivity to 5-FU,MTX,CDDP in the two gastric carcinoma cell lines. The IC50 of different chemotherapeutic agents in ASOON ＋ chemotherapy groups were significantly lower than those in chemotherapy groups. RT-PCR showed that tranfection with Cyclin D1 ASODN lead to a decrease in Cyclin D1 ,TS and DHFR mRNA and an increase in TP mRNA as determined by RT-PCR at 24 h ,the alterations were more significant at 48 h. Conclusion Cyclin D1 can enhance the chemosensitivity to multiple chemotherapeutic agents in gastric carcinoma cells ,and this effect maybe due to the altered expression of enzymes related to metabolism of chemotherapeutic agents.