BACKGROUND Substantial clinical and preclinical evidence have indicated the association between amide-linked local anesthesia and the long-term outcomes of cancer patients. However, the potential effects of local anesthesia on cancer recurrence are inconclusive and the underlying mechanisms remain poorly understood. METHODS We systematically examined the effects of three commonly used local anesthetics in melanoma cells and analyzed the underlying mechanisms focusing on small GTPases. RESULTS Ropivacaine and lidocaine but not bupivacaine inhibited migration and proliferation, and induced apoptosis in melanoma cells. In addition, ropivacaine and lidocaine but not bupivacaine significantly augmented the in vitro efficacy of vemurafenib (a B-Raf inhibitor for melanoma with BRAF V600E mutation) and dacarbazine (a chemotherapeutic drug). Mechanistically, ropivacaine but not bupivacaine decreased the activities of Ras superfamily members with the dominant inhibitory effects on RhoA and Ras, independent of sodium channel blockade. Rescue studies using constitutively active Ras and Rho activator calpeptin demonstrated that ropivacaine inhibited migration mainly through RhoA whereas growth and survival were mainly inhibited through Ras in melanoma cells. We further detected a global reduction of downstream signaling of Ras and RhoA in ropivacaine-treated melanoma cells. CONCLUSION Our study is the first to demonstrate the anti-melanoma activity of ropivacaine and lidocaine but not bupivacaine, via targeting small GTPases. Our findings provide preclinical evidence on how amide-linked local anesthetics could affect melanoma patients.

中文翻译：

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通过抑制Ras和RhoA信号传导而独立于钠通道阻滞，酰胺连接的局部麻醉药对黑素瘤细胞的细胞毒性。

背景技术大量的临床和临床前证据表明，酰胺连接的局部麻醉与癌症患者的长期预后之间存在关联。然而，局部麻醉对癌症复发的潜在影响尚无定论，其潜在机制仍知之甚少。方法我们系统地研究了三种常用的局部麻醉药在黑素瘤细胞中的作用，并分析了以小GTP酶为基础的潜在机制。结果罗哌卡因和利多卡因抑制布洛卡因的迁移和增殖，并诱导黑素瘤细胞凋亡。此外，罗哌卡因和利多卡因而不是布比卡因显着增强了维拉非尼（一种具有BRAF V600E突变的黑色素瘤的B-Raf抑制剂）和达卡巴嗪（一种化学治疗药物）的体外疗效。从机理上讲，罗哌卡因而不是布比卡因降低了Ras超家族成员的活性，对RhoA和Ras具有显着的抑制作用，而与钠通道阻滞无关。使用组成型活性Ras和Rho激活物calpeptin进行的拯救研究表明，罗哌卡因主要通过RhoA抑制迁移，而黑素瘤细胞中的生长和存活主要通过Ras抑制。我们进一步检测到罗哌卡因治疗的黑色素瘤细胞中Ras和RhoA下游信号的整体减少。结论我们的研究是第一个通过靶向小GTP酶证明罗哌卡因和利多卡因而不是布比卡因具有抗黑素瘤活性的研究。我们的发现为酰胺连接的局部麻醉药如何影响黑色素瘤患者提供了临床前证据。罗哌卡因而非布比卡因降低了Ras超家族成员的活性，对RhoA和Ras具有显着的抑制作用，而与钠通道阻滞无关。使用组成型活性Ras和Rho激活物calpeptin进行的拯救研究表明，罗哌卡因主要通过RhoA抑制迁移，而黑素瘤细胞中的生长和存活主要通过Ras抑制。我们进一步检测了罗哌卡因治疗的黑色素瘤细胞中Ras和RhoA下游信号的整体减少。结论我们的研究是第一个通过靶向小GTP酶证明罗哌卡因和利多卡因而不是布比卡因具有抗黑素瘤活性的研究。我们的发现为酰胺连接的局部麻醉药如何影响黑色素瘤患者提供了临床前证据。罗哌卡因而不是布比卡因降低了Ras超家族成员的活性，对RhoA和Ras具有显着的抑制作用，而与钠通道阻滞无关。使用组成型活性Ras和Rho激活物calpeptin进行的拯救研究表明，罗哌卡因主要通过RhoA抑制迁移，而黑素瘤细胞中的生长和存活主要通过Ras抑制。我们进一步检测了罗哌卡因治疗的黑色素瘤细胞中Ras和RhoA下游信号的整体减少。结论我们的研究是第一个通过靶向小GTP酶证明罗哌卡因和利多卡因而不是布比卡因具有抗黑素瘤活性的研究。我们的发现为酰胺连接的局部麻醉药如何影响黑色素瘤患者提供了临床前证据。