摘要
目的:为了深入了解十六烷基磷酸胆碱(HePC)在长期应用中是否产生耐药,通过长期诱导使KB细胞对HePC产生耐药,研究耐药株KBr的耐药特性,为其应用提供依据。方法:耐药株的建立采用低浓度HePC缓慢诱导,逐步增加浓度,直至KB细胞产生明显耐药;应用MTT法比较KB细胞与KBr细胞对不同化疗药物的IC50值;用RTPCR检测mdrl,MRP和GSTpi的mRNA;用免疫细胞化学方法检测P糖蛋白(Pgp)表达。结果:KB细胞经过HePC70周的长期诱导,产生耐药性,耐药倍数为32倍。KBr对长春新碱和秋水仙素有显著的交叉耐药,而对柔红霉素、5氟尿嘧啶、氮芥、卡氮芥、放线菌素D以及足叶乙甙(VP16)无明显的交叉耐药性。KBr细胞具mdrl基因和Pgp蛋白的阳性表达,MRP和GSTpi基因为阴性表达。结论:通过长期诱导KB细胞产生对HePC的耐药,耐药株对长春新碱和秋水仙素有交叉耐药,mdrl基因和Pgp高表达可能与耐药有关。
objective:
To induce tolerance to hexadecylphosphocholine(HePC) in human epidermoid tumour cell line
KB with gradually increasing concentration of HePC. To study the characteristic of the
resistance subline KBr cells. Methods: KB cells were adapted to HePC starting from low
concentration of HePC, the concentration of HePC was raised gradually until the KB cells had
obvious resistance to it.MTT assay was used for test of cross resistance to chemotherapeutics
agents in KBr cells.The expression of mdr1,mrp and GSTpi gene was determined by RTPCR.
Immunocytochemistry was used for expression of Pglycoprotein(Pgp).ZResults: KB cells were
adapted to HePC starting from a concentration at 004 mol/L. When the cells tolertated to HePC
and showed normal growth and high viability, the concentration of HePC was raised
accordingly.The adaptation was stopped when the concentration of HePC reached to 45 mol/L,
cells had obvious resistance to HePC. MTT assay showed the KBr cells had 32fold higher IC50
of HePC than parental KB cells.KBr cells also had significant resistance to vincristine and
colchicine although KBr cells were still sensitive to mechlorethamine,carmustine,actinomycin
D, etoposide,doxorubicin and 5fluorouracil.The IC50 of vincristine and colchicine on KBr cells
were respectively 1354fold and 44697fold much higher than that of KB cells. Our study showed
that Kbr cells had no expression of mrp and GSTpi gene, but there were mdr1 gene and Pgp
positive expression in resistant KBr cells. Conclusion: KB cells show obvious resistance to
HePC after long term induction.The resistant subline KBr cells have cross resistance to
vincristine and colchicine.The positive expression of mdr1 and Pgp in KBr cells might be
associated with the resistance.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
1999年第3期266-269,共4页
Chinese Journal of Cancer