Lnc46198半番鸭胚胎期特异性表达及对鸭胚成纤维细胞增殖和凋亡的影响

Specific expression of lnc46198in Mulard duck embryo and its impacts on cell proliferation and apoptosis of duck emybryo fibroblasts

为分析TCONS_00246198(简称lnc46198)在半番鸭胚胎期发育过程中的特异性表达及对鸭胚成纤维细胞(Duck embryo fibroblasts,DEF)增殖或凋亡的影响,本研究采用RT-qPCR分析lnc46198在10、15、20和25 d不同胚龄半番鸭性腺及半番鸭12.5 d胚龄大脑、心脏、肝脏、肺脏、肌肉、性腺和肾脏等不同组织中的表达情况。采用全基因合成法构建鸭胚成纤维细胞lnc46198-pcDNA3.1过表达载体,利用流式细胞术和CCK8法检测细胞增殖情况,采用Annexin V-FITC/PI流式和RT-qPCR检测细胞凋亡情况。结果表明:1)lnc46198在半番鸭各种组织和不同胚龄性腺中均表达,其在半番鸭12.5 d胚龄性腺的表达量极显著高于其他组织(P<0.01),且性腺中相对表达量随着胚龄增加逐渐升高,提示lnc46198可能在半番鸭性腺发育中发挥重要作用。2)鸭胚成纤维细胞lnc46198过表达组细胞活力与对照组相比极显著降低(P<0.01),表明过表达lnc46198可抑制鸭胚成纤维细胞的增殖活力。3)过表达组细胞凋亡率是pcDNA3.1组的7.30倍(P<0.01),表明过表达lnc46198可促进鸭胚成纤维细胞凋亡。综上,lnc46198在不同胚龄半番鸭性腺中动态表达,在半番鸭12.5胚龄性腺中特异表达,可抑制鸭胚成纤维细胞增殖,并促进鸭胚成纤维细胞凋亡,本研究揭示了lnc46198的特异性表达及在鸭细胞增殖或凋亡中的功能,可为半番鸭性腺发育差异机制提供科学参考依据。

The study was to analyze the expression of TCONS_00246198,also known as lnc46198,during embryonic development of Mulard duck and determine its impacts on the proliferation and apoptosis of duck embryonic fibroblasts.The RT-qPCR technique was employed to analyze the expression of lnc46198in various tissues including different embryo age such as 10,15,20and 25d,as well as the brain,heart,liver,lung,muscle,gonad,and kidney of Mulard duck in this study.The lnc46198-pcDNA3.1overexpression line of duck embryo fibroblasts was constructed by whole gene synthesis.Flow cytometry and CCK8assay were used to detect cell proliferation,while Annexin V-FITC flow and RT-PCR were used to detect apoptosis.The results showed that:1)The lnc46198was expressed that in all embryonic gonads,and its relative expression levels gradually increased with the age of the embryo,and the expression level of lnc46198in the gonads of Mulard duck at 12.5dembryo age was found to be significantly higher than that in all other tissues(P<0.01),suggesting potentially important role for lnc46198in the gonadal development of Mulard duck.2)Compared with blank group and pcDNA3.1group(control group),the cell viability of duck embryo fibroblasts in lnc46198overexpression group(test group)was significantly decreased(P<0.01),indicating that overexpression of lnc46198inhibited the proliferation viability of duck embryo fibroblasts.3)The overexpression group exhibited a significantly higher apoptosis rate(7.30times)of duck embryo fibroblasts compared with pcDNA3.1group(P<0.01),which has been suggested that the overexpression of lnc46198leads to an increase in apoptosis in duck embryo fibroblasts.In conclusion,the study found that lnc46198was dynamically expressed in the gonads at various embryonic stages,with specific expression in the gonads of Mulard ducks at 12.5embryo age.Furthermore,the results showed that overexpression of lnc46198inhibited the proliferation of duck embryo fibroblasts while promoting apoptosis.The study uncovered the distinct expression of lnc46198and its impact on the proliferation and apoptosis of duck cells,offering valuable scientific insight into the mechanism underlying the difference in gonad development in Mulard duck.