AKT途径在13-MTD诱导膀胱癌T24细胞凋亡中的作用研究

13-Methyltetradecanoic acid (13-MTD) is a saturated branched-chain fatty acid purified from soy fermentation products. 13-MTD has been first used to treat bladder cancer in our previous research through the mechanism of inducing apoptosis of T24 human bladder cancer cells. Our further study showed that this mechanism was through stress pathway via three major apoptotic pathways: Akt, JNK and P38. However, how these three pathways interact is not clear. Based on the current literature and our previous study, we propose that down-regulation of AKT plays a crucial role in inducing apoptosis of bladder cancer. Besides, JNK and P38 signaling pathways are activated by AKT, which will trigger apoptosis of cancer cells. This study will further reveal the interaction mechanisms of these three signaling pathways by using many methods such as RNA interference and Immunoprecipitation, seeking to find out whether down-regulation of AKT by 13-MTD could activate the JNK and p38 signaling pathways and whether this process is conducted though phosphorylation of Apoptosis Signal-regulating Kinase 1 (ASK1), and further to test the hypothesis that down-regulation of AKT induces the apoptosis of human bladder cancer cells. This research will provide scientific evidence to apply 13-MTD to human bladder cancer treatment.

13-甲基十四烷酸（13-methyltetradecanoic acid,13-MTD）是饱和十五碳脂肪酸，是大豆发酵的提取物。我们首次将其应用于膀胱癌的治疗研究中，已经证明了13-MTD可以诱导膀胱癌T24细胞凋亡，我们进一步的研究发现：13-MTD通过应激途径诱导膀胱癌细胞凋亡，并发现了三条起效的细胞凋亡途径：Akt、JNK、P38途径，但其相互作用均不清楚。基于相关的文献我们推测Akt的下调可能起了关键的作用，并能作用于JNK/P38途径，促进肿瘤细胞的凋亡。本项目拟通过RNA干扰、免疫沉淀等方法对该机制进行深入研究，明确13MTD下调Akt存活通道是否导致JNK/P38通道的激活，这一过程是否通过Akt磷酸化ASK1来实现，进而验证13-MTD通过下调Akt存活通道诱导肿瘤细胞凋亡的推断，为13-MTD用于肿瘤防治提供理论依据。

13-甲基十四烷酸（13-methyltetradecanoic acid,13-MTD）是饱和十五碳脂肪酸，是大豆发酵的提取物。我们首次将其应用于膀胱癌的治疗研究中，已经证明了13-MTD可以诱导膀胱癌T24细胞凋亡，我们进一步的研究发现：13-MTD通过应激途径诱导膀胱癌细胞凋亡，并发现了三条起效的细胞凋亡途径：Akt、JNK、P38途径，但其相互作用均不清楚。试验中我们采用模拟AKT2通路、阻断JNK/P38通路及上调AKT2通路发现：13MTD可以下调AKT2存活通道导致JNK/P38通道激活来诱导膀胱癌T24细胞凋亡。通过免疫共沉淀、RNA干扰等方法发现AKT2和JNK/P38通道之间是通过Akt磷酸化ASK1的Ser83位点而实现的。。