背景:头颈部鳞状细胞癌(HNSCC)是一种常见的癌症,五年生存率约为60%,表明需要新的治疗方法。BH3模拟物是抑制抗凋亡Bcl-2家族蛋白的小分子,导致细胞凋亡诱导。
方法:我们使用Myogel矩阵进行了高通量筛选,以鉴定辐照与12种HNSCC细胞系中新型BH3模拟物A-1155463,A-1331852和navitoclax之间的协同作用。正常(NOF)和癌症相关成纤维细胞(CAF),和发育不良的角质形成细胞(ODA)。接下来,我们在凋亡试验中检查了协同作用,然后在选定的HNSCC细胞系上进行克隆形成测定和Myogel球体。最后,我们应用斑马鱼幼虫异种移植来验证navitoclax和A-1331852的效果。
结果:所有三种BH3模拟物在八种HNSCC细胞系和ODA中表现出与辐射的强烈协同作用,但不在NOF和CAF中。A-1155463和A-1331852诱导细胞凋亡和减少增殖,和辐射一起,显著增加细胞凋亡和抑制增殖。与对照相比,A-1331852和navitoclax显着降低了克隆性,与单药治疗或放疗相比,联合治疗导致克隆源性降低。然而,与navitoclax或A-1155463不同,只有A-1331852能显著减少癌细胞的侵袭.此外,在球体和斑马鱼中,照射似乎无效,未能显着增加药物效果。在斑马鱼中,A-1331852和navitoclax可显着减少肿瘤面积和转移。
结论:我们的发现鼓励BH3模拟物的进一步临床前研究,特别是A-1331852,作为单一药物或与辐射联合治疗HNSCC。
BACKGROUND: Head and neck squamous cell carcinoma (HNSCC) is a common cancer with a five-year survival rate around 60%, indicating a need for new treatments. BH3 mimetics are small molecules that inhibit anti-apoptotic Bcl-2 family proteins, resulting in apoptosis induction.
METHODS: We performed a high-throughput screen using a Myogel matrix to identify the synergy between irradiation and the novel BH3 mimetics A-1155463, A-1331852, and
navitoclax in 12 HNSCC cell lines, normal (NOF) and cancer-associated fibroblasts (CAF), and dysplastic keratinocytes (ODA). Next, we examined synergy in an apoptosis assay, followed by a clonogenic assay and a Myogel spheroid on selected HNSCC cell lines. Finally, we applied zebrafish larvae xenograft to validate the effects of
navitoclax and A-1331852.
RESULTS: All three BH3 mimetics exhibited a strong synergy with irradiation in eight HNSCC cell lines and ODAs, but not in NOFs and CAFs. A-1155463 and A-1331852 induced apoptosis and reduced proliferation, and together with irradiation, significantly increased apoptosis and arrested proliferation. A-1331852 and
navitoclax significantly decreased the clonogenicity compared with the control, and combination treatment led to a decreased clonogenicity compared with monotherapy or irradiation. However, unlike
navitoclax or A-1155463, only A-1331852 significantly reduced cancer cell invasion. Furthermore, in spheroid and zebrafish, irradiation appeared ineffective and failed to significantly increase the drug effect. In the zebrafish, A-1331852 and
navitoclax significantly reduced the tumor area and metastasis.
CONCLUSIONS: Our findings encourage the further preclinical investigation of BH3 mimetics, particularly A-1331852, as a single agent or combined with irradiation as a treatment for HNSCC.