澳洲宝石鲈热休克蛋白90-β基因(HSP90-β)的cDNA克隆与表达

热休克蛋白(heat shock proteins,HSPs)在鱼类的应激与免疫反应中起着重要的作用,HSP90-β是该家族的重要成员。为了探讨澳洲宝石鲈(Scortum barcoo)HSP90-β在免疫应答中的作用,本研究首次克隆得到了澳洲宝石鲈HSP90-β的全长cDNA序列。该序列长2708 bp,其中5’UTR长86 bp,3’UTR长444 bp,ORF长2178 bp,编码725个氨基酸,相对分子质量约为83200,具有5个HSP90家族的特征序列,没有信号肽序列和跨膜结构域,与其他鱼类HSP90-β氨基酸的序列一致性皆超过90%。正常情况下,澳洲宝石鲈HSP90-βmRNA主要在肝脏表达。而感染无乳链球菌(Streptococcus agalactiae)后,其在肝组织中的表达呈先降低后增加的趋势,在脾脏、头肾和脑组织则呈先上升后下降的趋势。这些结果表明澳洲宝石鲈HSP90-β可能参与了无乳链球菌感染引发的免疫反应。

千里光热激蛋白90-3(Hsp90-3)的生物信息学与功能分析

Hsp90是真核细胞重要的一类分子伴侣,与植物的生长发育、抗逆性、信号转导及生物进化等功能密切相关。为了深入理解高等植物Hsp90结构与功能的关系,该研究从千里光(Senecio scandens)全长cDNA文库中分离到Hsp90-3基因。序列分析结果表明,该基因编码699个氨基酸的多肽,与拟南芥(Arabidopsis thaliana)AtHsp90-3(登录号:NP_200412.1)的同源性最高,为93.71%;预测蛋白质的分子量为79.78 kD,理论等电点为5.08。信号序列分析结果发现,该蛋白主要定位于细胞的细胞核、过氧化物酶体、叶绿体类囊体膜及叶绿体基质中,提示作为分子伴侣,高等植物Hsp90-3参与细胞内膜系统蛋白质的转运。结构与功能分析发现,该蛋白有3个结构域及1个连接区,推测Hsp90-3在真核细胞的信号转导、转录调控及胁迫表达等过程中发挥重要功能。

热休克蛋白90-α在胃癌诊断中的意义

目的了解热休克蛋白90-α(heatshockprotein90-α,HSP90-α)在胃癌患者外周血的表达情况,并分析其与胃癌患者临床病理学特点及营养风险之间的关系,探讨热休克蛋白90-α在胃癌诊断中的意义。方法选取2014年7月至2018年4月期间在烟台市烟台山医院接受手术并证实为胃癌的316例患者,术前采用酶联免疫法(ELISA)检测其血清中HSP90-α的水平。结果胃癌患者血清HSP90-α水平(119.15±65.47pg/ml)明显高于健康对照组(49.31±20.43pg/ml)及良性病变组(59.36±21.52pg/ml)(P<0.05)。此外,HSP90-α的表达与肿瘤的组织学分级、病理分期、肌肉减少症相关(P<0.05)。区别胃癌组与健康对照组及良性病变组的临界值分别是69.88pg/ml与72.90pg/ml,对应敏感度分别为85.4%和82.6%,特异性为85.7%和80.7%。结论HSP90-α在胃癌患者外周血水平升高与组织学分级差、病理分期晚、合并肌肉减少症有关,可能成为一种新的辅助诊断胃癌的肿瘤标志物。

Expression of heat shock protein 90 β in human gastric cancer tissue and SGC7901/VCR of MDR type gastric cancer cell line

Objective To examine the expression of heat shock protein (HSP) 90 β in human gastric cancer tissue and SGC7901/VCR of MDR type gastric cancer cell line Methods Immunohistochemical staining and in situ hybridization methods Results Heat shock protein 90 β was mainly located in the cell cytoplasma and weakly expressed in non cancerous gastric mucosa The expression rates of HSP90 β in normal gastric mucosa, gastritis and para cancer tissues were 11 76%, 13 04% and 11 42% respectively,and there were no significant differences between them ( P >0 05) The expression of HSP90 β was increased in gastric cancer The positive rate of HSP90 β in gastric cancer tissue was 30 00%, and was higher than non cancerous gastric mucosa ( P <0 05) The expression rates of HSP90 β in well differentiated, moderately differentiated, poorly differentiated gastric cancer and mucinous carcinoma were 15 38%, 31 25%, 33 33%, and 42 85% respectively The expression of HSP90 β in SGC7901/VCR of MDR type gastric cell line was higher than in its parental cell line SGC7901 In situ hybridization showed that the positive signal of HSP90 β was mainly located in the cell cytoplasma Conclusions The expression of HSP90 β was higher in gastric cancer tissue than in non cancerous gastric mucosa In gastric cancer tissue, the expression of HSP90 β was greater in poorly differentiated cancer tissue, and in SGC7901/VCR of MDR type gastric cancer cell line the expression of HSP90?β was higher than that in its parental cell line SGC7901

人参皂苷Rh_2(S)抑制HDAC6促白血病细胞凋亡作用研究

目的研究人参皂苷Rh2(S)促白血病K562和KG1a细胞凋亡的机制。方法 CCK-8法检测人参皂苷Rh2(S)对细胞增殖的影响;镜下观察细胞的生长情况;流式细胞术(FCM)检测细胞周期和凋亡;Western blotting法检测周期和凋亡相关蛋白以及组蛋白去乙酰化酶6(HDAC6)、热休克蛋白90(HSP90)、蛋白激酶B(Akt)、糖原合成酶激酶-3(GSK-3β)和β-连环蛋白(β-catenin)的表达。结果 CCK-8结果显示人参皂苷Rh2(S)对K562和KG1a细胞具有较明显的增殖抑制作用;FCM结果显示人参皂苷Rh2(S)能够将K562和KG1a细胞周期阻滞在G0/G1期,同时促进细胞凋亡;Westernblotting结果显示人参皂苷Rh2(S)能够抑制Bcl-2、Cyclin D1、HDAC6、HSP90、p-Akt和β-catenin蛋白的表达,促进Bax、Ac-α-tubulin和GSK-3β蛋白的表达。结论 Rh2(S)通过抑制HDAC6的表达导致HSP90的表达下降,进一步抑制Akt的活性,激活GSK-3β,最后抑制Wnt/β-catenin信号通路,促进白血病细胞凋亡。