Xie C-y, Kong J-r, Zhao C-s, Xiao Y-c, Peng T, Liu Y, Wang W-n
Key Laboratory of Ecology and Environmental Science in Guangdong Higher Education, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, College of Life Science, South China Normal University, Guangzhou 510631, PR China.
Key Laboratory of Ecology and Environmental Science in Guangdong Higher Education, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, College of Life Science, South China Normal University, Guangzhou 510631, PR China.
Dev Comp Immunol. 2016 Jun;59:77-88. doi: 10.1016/j.dci.2016.01.004. Epub 2016 Feb 4.
PTEN, a tumor suppressor gene, suppresses cell survival, growth, apoptosis, cell migration and DNA damage repair by inhibiting the PI3K/AKT signaling pathway. In this study, the full-length Litopenaeus vannamei PTEN (LvPTEN) cDNA was obtained, containing a 5'UTR of 59bp, an ORF of 1269bp and a 3'UTR of 146bp besides the poly (A) tail. The PTEN gene encoded a protein of 422 amino acids with an estimated molecular mass of 48.3 KDa and a predicted isoelectric point (pI) of 7.6. Subcellular localization analysis revealed that LvPTEN was distributed in both cytoplasm and nucleus, and the tissue distribution patterns showed that LvPTEN was ubiquitously expressed in all the examined tissues. Vibrio alginolyticus challenge induced upregulation of LvPTEN expression. Moreover, RNAi knock-down of LvPTEN in vivo significantly increased the expression of LvAKT mRNA, while reducing that of the downstream apoptosis genes LvP53 and LvCaspase3. LvPTEN knock-down also caused a sharp increase in cumulative mortality, bacterial numbers, and DNA damage in the hemolymph of L. vannamei following V. alginolyticus challenge, together with a sharp decrease in the total hemocyte count (THC). These results suggested that LvPTEN may participate in apoptosis via the PI3K/AKT signaling pathway in L. vannamei, and play an important role in shrimp innate immunity.
PTEN是一种肿瘤抑制基因,通过抑制PI3K/AKT信号通路来抑制细胞存活、生长、凋亡、细胞迁移和DNA损伤修复。在本研究中,获得了凡纳滨对虾PTEN(LvPTEN)的全长cDNA,除了poly(A)尾外,其包含59bp的5'UTR、1269bp的ORF和146bp的3'UTR。PTEN基因编码一个由422个氨基酸组成的蛋白质,估计分子量为48.3 kDa,预测等电点(pI)为7.6。亚细胞定位分析表明,LvPTEN分布于细胞质和细胞核中,组织分布模式显示LvPTEN在所有检测组织中均有广泛表达。溶藻弧菌攻击诱导LvPTEN表达上调。此外,体内RNA干扰敲低LvPTEN显著增加了LvAKT mRNA的表达,同时降低了下游凋亡基因LvP53和LvCaspase3的表达。敲低LvPTEN还导致凡纳滨对虾在溶藻弧菌攻击后血淋巴中的累积死亡率、细菌数量和DNA损伤急剧增加,同时总血细胞计数(THC)急剧下降。这些结果表明,LvPTEN可能通过PI3K/AKT信号通路参与凡纳滨对虾的凋亡,并在对虾先天免疫中发挥重要作用。