黑鲷催乳素基因PRL1和PRL2在急性盐胁迫环境下的表达分析

【目的】明确催乳素基因(PRL1和PRL2)在黑鲷不同组织及不同盐度胁迫下的表达模式,为黑鲷养殖过程中最适盐度范围的确定提供参考依据。【方法】通过RT-PCR扩增黑鲷PRL1和PRL2基因,以ExPASy、NetNGlyc及SWISS-MODEL等在线软件进行生物信息学分析,并采用实时荧光定量PCR检测PRL1和PRL2基因在黑鲷不同组织及不同盐度胁迫下的表达情况。【结果】黑鲷PRL1基因开放阅读框(ORF)全长639 bp,共编码212个氨基酸残基,其编码蛋白分子量为23.32 k D,理论等电点(pI)为6.70;黑鲷PRL2基因ORF全长750 bp,共编码249个氨基酸残基,其编码蛋白分子量为28.31 kD,pI为8.37。黑鲷PRL1和PRL2蛋白均由1个为信号肽结构域和1个为Hormone 1结构域组成;黑鲷PRL1蛋白二级结构中α-螺旋占69.34%、无规则卷曲占30.66%,包含1个糖基化位点及37个磷酸化位点;黑鲷PRL2蛋白二级结构中α-螺旋占67.07%、无规则卷曲占32.93%,包含1个糖基化位点及21个磷酸化位点。黑鲷PRL1和PRL2基因在12个待检测组织中均有表达,且均以脑组织中的相对表达量最高,显著高于其他组织中的相对表达量(P<0.05,下同);在盐度5‰处理组中,黑鲷PRL1基因在胁迫4 h开始显著上调,黑鲷PRL2基因在胁迫8 h开始极显著上调(P<0.01),二者均在胁迫24 h时达峰值,随后开始缓慢下降,至胁迫72 h时其相对表达量仍显著高于对照组;在盐度25‰和35‰处理组中,黑鲷PRL1和PRL2基因在胁迫72 h时其相对表达量均显著低于对照组。【结论】低盐胁迫能促进黑鲷PRL1和PRL2基因的表达,高盐胁迫则抑制黑鲷PRL1和PRL2基因的表达,即黑鲷PRL1和PRL2基因在其适应淡水环境的过程中发挥重要调节作用。

Detection of PRL-2 gene expression in hepatocellular carcinoma by real-time fluorescence quantitative PCR

Objective:To quantitatively detect the expression level of PRL-2 in primary hepatocellular carcinoma using real-time fluorescence quantitative PCR.Methods:Total RNA isolated from human HCC and liver tissue adjacent to the tumor was reversely transcribed into cDNA.Real-time fluorescence quantitative PCR(Q-PCR) method was used to analyze the expres-sion level of PRL-2 gene.Results:The Q-PCR method was performed successfully to precisely detect RNA level.PRL-2 was expressed in all portal vein tumor thrombosis(PVTT) and HCC,but only in some paratumor tissue.The highest expression level of PRL-2 gene was recorded in PVTT;meanwhile expression level of PRL-2 was higher than that in paratumor liver tis-sues and in HCC(P < 0.01),and it was higher in HCC than that in paratumor liver tissues.Conclusion:The Q-PCR may be the most precise method to quantitatively detect RNA level and can be used in gene expression changes.The PRL-2 gene has higher expression in PVTT than that in HCC and in paratumor liver tissue cells,indicating that it plays an important role in the development and metastasis of the HCC.

绝经期妇女血清生殖激素水平的观察

目的探讨绝经期妇女血清生殖激素水平与年龄的关系,为绝经期妇女自我保健提供科学依据。方法选择绝经期妇女,根据年龄、绝经年限进行分组:自然绝经期组120例,人工绝经组40例,同时选择100例月经正常者作为对照组。结果自然绝经组和人工绝经组与对照组比较PRL、E2水平是下降的,FSH、LH水平是上升的;绝经期妇女随着年龄的增长,PRL、E2水平呈下降趋势,FSH、LH水平呈上升趋势。结论绝经期妇女PRL、E2水平与年龄成负相关,FSH、LH水平与年龄呈正相关。