Clusterin在Anip973/CDDP细胞株中的表达及其意义

Expression and Significance of Clusterin in Anip973/CDDP Cell Lines

背景与目的聚集素(Clusterin,CLU)在肿瘤发生发展过程中的作用及变化特点是国内外近年来关注的一个热点,但有关CLU在肺癌中表达变化与抗癌药物敏感性机制的研究目前仍无相关报道。本研究比较CLU在Anip973与Anip973/CDDP两种细胞系中的表达,探讨CLU与非小细胞肺癌耐药关系。方法采用Western blot从蛋白水平比较CLU在Anip973和Anip973/CDDP中的差异表达,并通过流式细胞仪检测CLU与p53、Bax之间的关系。结果在蛋白水平,CLU在Anip973/CDDP中的表达都明显高于Anip973(P<0.01);p53在Anip973/CDDP中的表达明显高于Anip973(P<0.01);bax在Anip973/CDDP中的表达与Anip973相比无统计学意义(P>0.05)。进入G0期-G1期的Anip973/CDDP耐药细胞明显多于非耐药细胞,而进入S期、G2期-M期的Anip973/CDDP耐药细胞明显少于非耐药细胞。结论CLU与NSCLC的CDDP耐药性相关;CLU、p53在Anip973/CDDP中呈异常表达,提示CLU基因和p53基因可能共同参与非小细胞肺癌耐药的发生。

Background and objective Clusterin （CLU） is a multifunctional protein which attracts much attentions in recent years due to its function and variation in tumor formation and development. So far, there have no reports on its variation in lung cancer and its relationship with sensitivity of anti-cancer drugs. This study was to compare the expression of clusterin in wild Anip973 and Anip973/CDDP, thus to investigate the correlation of clusterin expression to tumorigenesis, tumor development and drug resistance in non-small cell lung cancer （NSCLC）. Methods Western blot and RT-PCRwere used to compare the clusterin expression at protein level and mRNA level in Anip973/CDDP with that in wild Anip973. Flow cytometry was employed to detect and analyze the relationship between clusterin and cell cycle, pS3 or Bax. Results Clusterin protein expression levels were significantly higher in those Anip973/CDDP than those in wild Anip973. The pS3 expression levels were higher in Anip973/ CDDP than in Anip973 （P〈0.01）. The expression of Bax showed no significant difference between Anip973 and Anip973/CDDP （P〉0.05）. As to drug resistant lung cancer cell lines, Anip973/CDDP had more cells in G0-G1 stage while less cells in S stage or G2-M stage than non-resistant cell line. Conclusion Clusterin is associated with resistance to CDDP in NSCLC. The abnormal expression of dusterin and pS3 in Anip973/CDDP indicated that both clusterin and pS3 might be involved in drug resistance in NSCLC.