PI3K/Akt/MMP-9通路在枸橼酸芬太尼抑制胃癌侵袭转移中的作用

Fentanyl is a representative drug of opioids which is widely used in patients with cancer pain. However, it’s not clear whether fentanyl affects the progression of cancer. In our previous study, we found that fentanyl inhibited growth, proliferation, invasion and metastasis of gastric cancer cells and downregulated pAkt and MMP-9 expression. In order to analyze the roles of PI3K/Akt/MMP-9 pathway on fentanyl inhibiting the progression of gastric cancer, we first test interacting of PI3K, Akt and MMP-9 by GST pull-down method. Moreover, we construct MMP-9 vectors and transfect these vectors into gastric cancer cells, or establish subcutaneous tumor model of gastric cancer in nude mice to investigate the effects of fentanyl or PI3K inhibitor on growth, proliferation, invasion, metastasis, angiogenesis and PI3K/Akt/MMP-9 pathway-related genes expression of gastric cancer in vitro and in vivo. At last, we observe the survival rate, the recurrence and metastasis of gastric cancer and blood MMP-9 expression in patients with cancer pain who were treated with fentanyl or other drugs in order to analyze the factors affecting survival time of patients and to elucidate the roles of PI3K/Akt/MMP-9 pathway on fentanyl inhibiting the invasion and metastasis of gastric cancer.

芬太尼是阿片类药物的代表，在癌痛治疗中应用广泛，但它对于癌症进展的影响还不清楚。我们的前期研究发现，枸橼酸芬太尼能够抑制胃癌细胞的生长增殖、侵袭转移，并下调细胞内pAkt、MMP-9的表达。为了进一步分析PI3K/Akt/MMP-9通路在芬太尼抑制胃癌进展中的作用，本项目首先通过GST pull-down实验验证PI3K、Akt和MMP-9的相互作用；然后通过构建MMP-9载体转染胃癌细胞或建立裸鼠人胃癌皮下瘤模型，在体外或体内观察芬太尼、PI3K抑制剂对胃癌生长增殖、侵袭转移、血管生成以及PI3K/Akt/MMP-9通路相关基因表达的影响；最后通过观察应用芬太尼等药物治疗癌痛的胃癌病人的存活率、复发转移情况及血MMP-9等因子的表达，对病人生存状况的相关因素进行分析，以深入探讨PI3K/Akt/MMP-9通路在芬太尼抑制胃癌侵袭转移中的作用。

枸橼酸芬太尼是阿片类镇痛药的代表性药物，在晚期癌痛病人中应用广泛，但芬太尼是抑制肿瘤侵袭转移还是促进肿瘤侵袭转移，现今报道不一。本项目通过建立裸鼠人胃癌皮下瘤动物模型，应用透射电镜、免疫组化、RT-PCR以及Western blot等技术观察到芬太尼抑制裸鼠人胃癌皮下瘤的生长、破坏肿瘤细胞的结构，NF-κB、Bcl-2、VEGF-A和MMP-9的表达降低，Bax的表达增加；体外细胞学实验中，分别以芬太尼、PI3K抑制剂或MMP-9抑制剂孵育胃癌MGC-803细胞，应用CCK-8法、克隆形成实验、流式细胞仪、划痕实验、透射电镜、RT-PCR以及Western blot等方法发现芬太尼抑制胃癌细胞的生长增殖、侵袭转移，促进胃癌细胞的凋亡，其机制与抑制PI3K/Akt/MMP-9通路有关；临床研究中，我们收集了510例胃癌并发癌痛的患者资料，发现性别、文化程度、病情了解情况、手术治疗、化疗、出现疼痛的时间、疼痛治疗史、NRS评分和KPS评分是胃癌患者预后的独立影响因素。总之，本研究揭示了芬太尼抑制胃癌侵袭转移的分子机制，从而为芬太尼在癌痛治疗中的应用提供指导。