Repurposing the serotonin agonist Tegaserod as an anticancer agent in melanoma: molecular mechanisms and clinical implications.
重新利用 5-羟色胺激动剂替加色罗作为黑色素瘤的抗癌药物: 分子机制和临床意义。
- 作者列表："Liu W","Stachura P","Xu HC","Umesh Ganesh N","Cox F","Wang R","Lang KS","Gopalakrishnan J","Häussinger D","Homey B","Lang PA","Pandyra AA
BACKGROUND:New therapies are urgently needed in melanoma particularly in late-stage patients not responsive to immunotherapies and kinase inhibitors. METHODS:Drug screening, IC50 determinations as well as synergy assays were detected by the MTT assay. Apoptosis using Annexin V and 7AAD staining was assessed using flow cytometry. TUNEL staining was performed using immunocytochemistry. Changes in phosphorylation of key molecules in PI3K/Akt/mTOR and other relevant pathways were detected by western blot as well as immunocytochemistry. To assess in vivo anti-tumor activity of Tegaserod, syngeneic intravenous and subcutaneous melanoma xenografts were used. Immunocytochemical staining was performed to detect expression of active Caspase-3, cleaved Caspase 8 and p-S6 in tumors. Evaluation of immune infiltrates was carried out by flow cytometry. RESULTS:Using a screen of 770 pharmacologically active and/or FDA approved drugs, we identified Tegaserod (Zelnorm, Zelmac) as a compound with novel anti-cancer activity which induced apoptosis in murine and human malignant melanoma cell lines. Tegaserod (TM) is a serotonin receptor 4 agonist (HTR4) used in the treatment of irritable bowel syndrome (IBS). TM's anti-melanoma apoptosis-inducing effects were uncoupled from serotonin signaling and attributed to PI3K/Akt/mTOR signaling inhibition. Specifically, TM blunted S6 phosphorylation in both BRAFV600E and BRAF wildtype (WT) melanoma cell lines. TM decreased tumor growth and metastases as well as increased survival in an in vivo syngeneic immune-competent model. In vivo, TM also caused tumor cell apoptosis, blunted PI3K/Akt/mTOR signaling and decreased S6 phosphorylation. Furthermore TM decreased the infiltration of immune suppressive regulatory CD4+CD25+ T cells and FOXP3 and ROR-γt positive CD4+ T cells. Importantly, TM synergized with Vemurafenib, the standard of care drug used in patients with late stage disease harboring the BRAFV600E mutation and could be additively or synergistically combined with Cobimetinib in both BRAFV600E and BRAF WT melanoma cell lines in inducing anti-cancer effects. CONCLUSION:Taken together, we have identified a drug with anti-melanoma activity in vitro and in vivo that has the potential to be combined with the standard of care agent Vemurafenib and Cobimetinib in both BRAFV600E and BRAF WT melanoma.
背景: 黑色素瘤迫切需要新的治疗方法，尤其是对免疫疗法和激酶抑制剂无反应的晚期患者。 方法: 采用 MTT 法检测药物筛选、 IC50 测定及协同作用试验。使用 Annexin V 和 7AAD 染色的细胞凋亡使用流式细胞术进行评估。使用免疫细胞化学进行 TUNEL 染色。Western blot 和免疫细胞化学检测 PI3K/Akt/mTOR 等相关通路中关键分子磷酸化的变化。为了评估替加色罗的体内抗肿瘤活性，使用同基因静脉和皮下黑色素瘤异种移植物。免疫细胞化学染色检测肿瘤中活性 Caspase-3 、 cleaved Caspase 8 和 p-S6 的表达。通过流式细胞术进行免疫浸润的评价。 结果: 使用 770 个药理学活性和/或 FDA 批准的药物的筛选，我们确定了替加色罗 (Zelnorm，Zelmac) 作为一种具有新型抗癌活性的化合物，诱导小鼠和人恶性黑色素瘤细胞系凋亡。替加色罗 (TM) 是一种 5-羟色胺受体 4 激动剂 (HTR4)，用于治疗肠易激综合征 (IBS)。TM 的抗黑色素瘤凋亡诱导作用与 5-羟色胺信号解偶联，归因于 PI3K/Akt/mTOR 信号抑制。具体而言，TM 在 BRAFV600E 和 BRAF 野生型 (WT) 黑色素瘤细胞系中均钝化了 S6 磷酸化。TM 减少了体内同基因免疫活性模型中的肿瘤生长和转移，并增加了生存率。在体内，TM 还引起肿瘤细胞凋亡，PI3K/Akt/mTOR 信号减弱，S6 磷酸化降低。此外，TM 降低了免疫抑制性调节性 CD4 + CD25 + T 细胞和 FOXP3 和 ROR-γ T 阳性 CD4 + T 细胞的浸润。重要的是，TM 与 Vemurafenib 协同作用, 在携带 BRAFV600E 突变的晚期疾病患者中使用的标准护理药物，并且可以在 BRAFV600E 和 BRAF WT 黑色素瘤细胞系中与 Cobimetinib 相加或协同组合诱导抗癌作用。。 结论: 综合起来看, 我们已经确定了一种具有体外和体内抗黑色素瘤活性的药物，该药物有可能与标准护理剂 Vemurafenib 和 Cobimetinib 在 BRAFV600E 和 BRAF WT 黑色素瘤中联合使用。
METHODS::In colorectal cancer (CRC), hepatic arterial infusion (HAI) chemotherapy may convert primarily unresectable CRC liver metastases (CLM) into resectability, although the risk of metastatic recurrence remains high after CLM ablation. We investigated the role of antitumour immunity invoked by first-line oxaliplatin-HAI for long-term CLM outcome. In a prospective study cohort of primarily unresectable CLM, we assessed patients' fms-related tyrosine kinase 3 ligand (FLT3LG) in serum, reflecting opportune intratumoural immune activity, at baseline and following 1-3 sequences of oxaliplatin-HAI. The end points were CLM resectability and overall survival. Patients who presented an immediate twofold increment of circulating FLT3LG during the treatment and at its completion were scored as CLM resectable (16.4% with both features), were alive at final follow-up 8-12 years later. All patients experienced FLT3LG increase during the treatment course, but those who remained unresectable or had the disease converted but presented a slow and gradual FLT3LG accretion, later died of the metastatic disease. These data provide further support to our previous findings that tumour-directed immunity invoked by oxaliplatin-containing therapy predicts excellent outcome of early advanced CRC if macroscopic tumour ablation is rendered possible by the 'classic' tumour response to the cytotoxic treatment.
METHODS::Prostate cancer is one of the primary causes of death around the world. As an important drug, flutamide has been used in the clinical diagnosis of prostate cancer. However, the over dosage and improper discharge of flutamide could affect the living organism. Thus, it necessary to develop the sensor for detection of flutamide with highly sensitivity. In this paper, we report the synthesis of lanthanum cobaltite decorated halloysite nanotube (LCO/HNT) nanocomposite prepared by a facile method and evaluated for selective reduction of flutamide. The as-prepared LCO/HNT nanocomposite shows the best catalytic performance towards detection of flutamide, when compared to other bare and modified electrodes. The good electrochemical performance of the LCO/HNT nanocomposite modified electrode is ascribed to abundant active sites, large specific surface area and their synergetic effects. Furthermore, the LCO/HNT modified electrode exhibits low detection limit (0.002 μM), wide working range (0.009-145 μM) and excellent selectivity with remarkable stability. Meaningfully, the developed electrochemical sensor was applied in real environmental samples with an acceptable recovery range.
METHODS::Several studies have indicated that cancer-associated fibroblasts (CAFs) could promote cancer progression in many malignancies. However, the mechanism by which CAFs promote the growth and metastasis of lung cancer remains poorly defined. In the present study, CAFs and normal fibroblasts (NFs) were isolated from human lung cancer and adjacent tissue. The data showed that the conditional medium (CM) of CAFs could increase the proliferation, migration and invasion of lung cancer cells. Vascular cell adhesion molecule-1 (VCAM-1) showed a higher expression in CAF-CM than NF-CM, and blocking VCAM-1 in CAF-CM attenuated the proliferation and invasion of cancer cells. Further, the results showed that VCAM-1 secreted from CAFs activated AKT and MAPK signaling via receptor α4β1 integrin (very-late antigen (VLA)-4) in lung cancer cells. Moreover, CAFs promoted VCAM-1 expression and tumor growth in vivo. Additionally, bioinformatics analysis indicated a positive correlation on the CAF marker protein alpha-smooth muscle actin (α-SMA) and VCAM-1 expression, which was associated with a poor prognosis in lung cancer patients. These findings demonstrate that the VCAM-1 secreted from CAFs enhances growth and invasion by activating the AKT and MAPK signaling of lung cancer cells.