组蛋白HIST1H2AC和HIST1H2BC基因在胰腺癌耐药细胞株中的表达

组蛋白HIST1H2AC和HIST1H2BC基因在胰腺癌耐药细胞株中的表达

（作者:___________单位: ___________邮编: ___________）

【摘要】目的:研究组蛋白HIST1H2AC和HIST1H2BC在培美曲塞诱导的胰腺癌耐药细胞株Puta8988中的基因和蛋白表达水平，以明确HIST1H2AC和HIST1H2BC基因与胰腺癌耐药的关系。方法:以两种剂量的培美曲塞诱导胰腺癌细胞株Puta8988，使其形成两个等级的获得性耐药细胞株Puta8988+、Puta8988++。采用qRT PCR和Western blotting方法分别检测HIST1H2AC和HIST1H2BC在耐药细胞株及其亲本(对照)中的基因和蛋白表达水平。结果:qRT PCR检测结果显示，组蛋白HIST1H2AC和HIST1H2BC基因在胰腺癌耐药株Puta8988+和Puta8988++中的表达水平显著上调(P0.05);Western blotting检测结果显示，组蛋白HIST1H2AC和HIST1H2BC在胰腺癌耐药株Puta8988+和Puta8988++中的蛋白表达水平也显著上调(P0.05)。结论：胰腺癌细胞株经培美曲塞诱导后，其组蛋白HIST1H2AC 和HIST1H2BC在基因和蛋白表达水平上均出现上调，提示HIST1H2AC 和HIST1H2BC基因可能与耐药有关。

【关键词】 HIST1H2AC; HIST1H2BC; 胰腺癌; Puta8988; 培美曲塞;

表达

[Abstract] Objective: To investigate the expression of HIST1H2AC and HIST1H2BC genes in pancreatic cancer cell line Puta8988 treated with pemetrexed, and to verify the relationship between the genes and pemetrexed resistance. Methods: Pancreatic cancer cell line Puta8988 was treated with two doses of pemetrexed to get Puta8988+ and Puta8988++ with pemetrexed resistance, and the expression levels of HIST1H2AC and HIST1H2BC and their genes in Puta8988, Puta8988+ and Puta8988++ were detected by qRT PCR and Western blotting respectively. Results: Results detected by qRT PCR showed that expression levels of HIST1H2AC and HIST1H2BC genes were significantly up regulated in Puta8988+ and Puta8988++(P0.05). Results detected by Western blotting showed that expression levels of HIST1H2AC and HIST1H2BC were also significantly up regulated in those cell lines with pemetrexed resistance(P0.05). Conclusion: Expressions of HIST1H2AC and HIST1H2BC and their genes in pancreatic cancer cell line Puta8988 treated with pemetrexed are significantly up regulated, which implies that these genes possibly plays a role in pemetrexed resistance of pancreatic cancer cell lines.

[Key words] HIST1H2AC; HIST1H2BC; pancreatic cancer; Puta8988; pemetrexed; expression

胰腺癌起病隐匿，手术切除率低，对传统放化疗不敏感，死亡发病比达98%，5年生存率6%[1-2]，是威胁人类健康的“隐秘杀手”。近年来的研究表明，辅助化疗有比传统化疗更好的疗效，已被认为是与手术治疗同样重要的治疗方法。与此同时，多种针对胰腺癌的靶向药物也在不断被发现。然而，耐药现象的存在仍然是导致辅助化疗效果欠佳或失败的主要原因。揭示胰腺癌化疗的耐药机制，建立更有效的化疗方法已成为国内外研究的重点[3];而从分子生物学的层面来研究和寻找耐药基因，然后给予必要的干预，以提高辅助化疗疗效的研究已成为当前最为关注的热点[4]。已有的研究表明，胰腺癌耐药性与MDR1(multidrug resistance1)、MRP(multidrug resistance associated protein gene)、LRP(lung resis 2 tance protein)、Bcl2(B cell lym phoma/leuke2)以及P53等基因有关，而这些基因的一个共同特点是与其他癌症的耐药性也有关[5]。这就提示我们可以借鉴其他癌症发现的耐药基因研究胰腺癌的耐药机制。有报道指出，组蛋白HIST1H2AC(Histone 1,H2ac)和HIST1H2BC(Histone 1,H2bc)基因与乳腺癌耐药性密切相关[6]。然而，它们是否在胰腺癌耐药细胞株中表达至今很少有报道。本实验针对胰腺癌耐药细胞株中HIST1H2AC和HIST1H2BC在基因和蛋白水平上的表达情况开展研究，以探讨它们与胰腺癌耐药的关系及其可能的分子机制，为研发对抗和逆转耐药的新方法提供新的靶点。