Yu Lan, Guo Qing-Ming, Wang Yu, Xu Yan, Liu Li, Zhang Xiao-Tao
Department of Stereotactic Radiotherapy, Affiliated Qingdao Central Hospital, Qingdao University, Qingdao, China.
Biotherapy Center, Affiliated Qingdao Central Hospital, Qingdao University, Qingdao, China.
J Thorac Dis. 2021 Apr;13(4):2404-2413. doi: 10.21037/jtd-21-442.
Recurrence of esophageal cancer (EC) after chemotherapy may mainly be explained by the existence of chemotherapy-resistant cells, and an effective drug against chemotherapy-resistant cells is highly sought. The aim of this study was to investigate the cytotoxicity of bispecific antibody solitomab combined with γ δ T cells on Eca109 cell spheres.
We cultured Eca109 cell spheres in serum-free medium, and the morphological differences between wild-type Eca109 cells and Eca109 cell spheres were compared by microscope and flow cytometry. Different concentrations of nanoparticle albumin-bound paclitaxel (Nab-PTX) and cisplatin were used to treat the two groups of cells and compare their drug resistance. Flow cytometry was then used to detect the expression level of epithelial cell adhesion molecule (EpCAM) and the cytotoxicity of γ δ T cells combined with bispecific antibody solitomab on the two groups.
Flow cytometry analysis showed that Eca109 cell spheres were smaller in size and had less cytoplasmic granules and CCK-8 assay showed that the viability of Eca109 cell spheres treated with different concentrations of Nab-PTX and cisplatin was significantly higher than that of wild-type Eca109 cells (P<0.05). Flow cytometry also showed that the expression level of EpCAM on Eca109 cell spheres was higher than that of wild-type Eca109 cells. Co-culture experiment showed that there was no significant difference in the cytotoxicity of γ δ T cells to wild-type Eca109 cells and Eca109 cell spheres without solitomab. However, after adding solitomab, the cytotoxicity of γ δ T cells to Eca109 cell spheres was significantly higher than that of wild-type Eca109 cells (P<0.05).
EC Eca109 cell spheres have strong stem cell characteristics such as multidrug resistance and may contain a high proportion of EC stem cells. Further, EC Eca109 cell spheres have a high expression level of EpCAM, and EpCAM may be one of the markers of EC stem cells. Therefore, EpCAM could be used as a potential molecular target of immunotherapy for EC, and solitomab may become an effective immunotherapeutic drug for chemotherapy-resistant EC cells.
食管癌(EC)化疗后复发可能主要归因于化疗耐药细胞的存在,因此人们迫切需要一种针对化疗耐药细胞的有效药物。本研究旨在探讨双特异性抗体索利托单抗联合γδT细胞对Eca109细胞球的细胞毒性。
我们在无血清培养基中培养Eca109细胞球,通过显微镜和流式细胞术比较野生型Eca109细胞与Eca109细胞球的形态差异。使用不同浓度的纳米白蛋白结合紫杉醇(Nab-PTX)和顺铂处理两组细胞,并比较它们的耐药性。然后采用流式细胞术检测上皮细胞黏附分子(EpCAM)的表达水平以及γδT细胞联合双特异性抗体索利托单抗对两组细胞的细胞毒性。
流式细胞术分析显示,Eca109细胞球体积较小,细胞质颗粒较少,CCK-8检测显示,用不同浓度的Nab-PTX和顺铂处理的Eca109细胞球的活力显著高于野生型Eca109细胞(P<0.05)。流式细胞术还显示,Eca109细胞球上EpCAM的表达水平高于野生型Eca109细胞。共培养实验表明,在没有索利托单抗的情况下,γδT细胞对野生型Eca109细胞和Eca109细胞球的细胞毒性没有显著差异。然而,加入索利托单抗后,γδT细胞对Eca109细胞球的细胞毒性显著高于野生型Eca109细胞(P<0.05)。
EC Eca109细胞球具有多药耐药等强大的干细胞特性,可能含有高比例的EC干细胞。此外,EC Eca109细胞球EpCAM表达水平较高,EpCAM可能是EC干细胞的标志物之一。因此,EpCAM可作为EC免疫治疗的潜在分子靶点,索利托单抗可能成为治疗化疗耐药EC细胞的有效免疫治疗药物。