摘要
目的探明YES相关蛋白(Yes-associated protein,YAP)如何通过调控miR-29a的表达来调节PTEN表达量。方法(1)以N2a细胞为研究对象,高表达及抑制YAP,测量其miR-29家族的表达量,并筛选出改变最明显的一个miRNA,并检测PTEN蛋白的表达;(2)通过改变miR-29a及YAP的蛋白水平,来检测YAP是通过何途径来调控PTEN蛋白的表达。结果 (1)通过转染了过表达YAP质粒后,RT-PCR的结果显示miR-29家族中的3个成员miR-29a,miR-29b,miR-29c都明显上调了。其中miR-29a上调了明显。而同时抑制了YAP的细胞中,miR-29的表达量都明显下降。说明YAP与miR-29的表达量之间是一种正相关的联系。另外一方面,PTEN的表达量则正好与miR-29及YAP相反,在抑制了YAP后,PTEN的表达量呈现的上升趋势,而过表达了YAP后,PTEN则明显下降。这说明YAP与PTEN之间是一种负相关的联系。(2)无论YAP的量改变如何,起到决定性作用的仍然还是miR-29a。下游的PTEN受YAP及miR-29a调控,而其中miR-29a作为作用中枢又受到上游YAP的调控。这些证据都揭示了YAP对PTEN的调节作用是通过对miR-29a的调节来实现的。结论该研究初步证实YAP能够通过对miR-29a的调控来实现对PTEN的表达,从而为治疗神经脊髓损伤提供研究基础。
Objective To explore how YES （Yes-associated protein） regulates the expression of PTEN by regulating the expression of miR-29a. Methods 1.N2a cells as the study object, high expression and inhibition of YAP, measuring the expression of the miR-29 family, and screening out the most obvious change in a miRNA, and detection of PTEN protein expression; 2.by changing the miR- Protein levels of 29a and YAP to detect the pathway by which YAP regulates PTEN protein expression. Results 1.RT-PCR showed that miR-29a, miR-29b and miR-29c were significantly up-regulated in the miR-29 family. Among them, miR-29a was significantly upregulated. In the cells that inhibited YAP at the same time, the expression of miR-29 decreased significantly. There was a positive correlation between the expression of YAP and miR-29. On the other hand, the expression level of PTEN was exactly the same as that of miR-29 and YAP. After inhibiting YAP, the expression level of PTEN showed an upward trend, while after overexpression of YAP, PTEN significantly decreased. This showed that there was a negative correlation between YAP and PTEN. 2.Regardless of how the YAP changes, the miR- 29a still played a decisive role. Downstream PTEN was regulated by YAP and milR-29a, and miR-29a was centrally regulated by upstream YAP. These evidences had revealed that the regulation of PTEN by YAP was achieved through the regulation of miR-29a. Conclusion This study initially confirmed that YAP can achieve the expression of PTEN through the regulation of miR-29a, which provides a basis for the treatment of neuronal spinal cord injury.
作者
孟甦
钱邦磊
余常麟
MENG Su;QIAN Bang-lei;YU Chang-lin(Department of General Surgery,the Second Affiliated Hospital of Soochow University,Suzhou,Jiangsu Province,215000 China;Department of Planning,Development and Performance Assessment,the Second Affiliated Hospital of Soochow U-niversity,Suzhou,Jiangsu Province,215000 China;Department of Orthopedics,Changzhou Second People＇s Hospital,Changzhou,Jiangsu Province,213164 China)
出处
《中外医疗》
2018年第25期21-23,共3页
China & Foreign Medical Treatment
