Institute of Anatomy and Cell Biology, School of Medicine, National Yang Ming University, Taipei, Taiwan, ROC.
General Surgery Division, Far Eastern Memorial Hospital, New Taipei City, Taiwan, ROC.
Asian Pac J Cancer Prev. 2023 Apr 1;24(4):1297-1305. doi: 10.31557/APJCP.2023.24.4.1297.
Human umbilical cord mesenchymal stem cells (hUCMSCs) have high therapeutic value in cancer treatment. We have found that pre-activating hUCMSCs with IL-1β promotes tumor necrosis factor-related apoptosis inducing ligand (TRAIL) expression and facilitates anti-tumor effect. Furthermore, embelin has been found to induce apoptosis of different cancer cell lines by upregulating the expression of TRAIL receptor 1 (DR4) and TRAIL receptor 2 (DR5). This study investigated whether IL-1β induced TRAIL-expressing hUCMSCs, in combination with low-dose embelin, could further induce apoptosis in breast cancer cell lines.
MTT assay was used to examine the cytotoxicity of embelin in MDA-MB-231 and MCF-7. To detect the interested protein expression in cells, Western blot and cell immunofluorescence were used to double-confirm the observed results. Annexin V/PI apoptosis assay was detected by flow cytometry to analyze the apoptosis rate of embelin treated breast cancer cell lines and the effect of co-culturing with breast cancer cells and hUCMSCs.
Using Western blot and immunofluorescence, we found that breast cancer cell lines treated with low-dose embelin (2.5-5 μM) increased the expression of apoptosis-related receptor DR4, DR5 and the cleaved caspase 8, 9 and 3. Moreover, TRAIL expression was enhanced in IL-1β induced hUCMSCs. Combining these observations, we expected that coculturing IL-1β induced hUCMSCs with low dose embelin treated MDA-MB-231 and MCF-7 cells might enhance the apoptosis of breast cancer cells. We confirmed via flow cytometry that coculture of IL-1β induced TRAIL-expressing hUCMSCs and embelin treated MDA-MB-231 and MCF-7 cells enhances the apoptosis rate of these breast cancer cells.
We found that embelin upregulated the expression of DR4 and DR5 to increase the TRAIL-mediated apoptosis in breast cancer cell lines. Low dose embelin treated breast cancer cell lines in combination with IL-1β induced TRAIL-expressing hUCMSCs may become a potential anti-tumor therapy.
人脐带间充质干细胞（hUCMSCs）在癌症治疗中有很高的治疗价值。我们发现，用白细胞介素-1β（IL-1β）预先激活 hUCMSCs 可以促进肿瘤坏死因子相关凋亡诱导配体（TRAIL）的表达，并促进抗肿瘤作用。此外，研究发现，埃博霉素通过上调 TRAIL 受体 1（DR4）和 TRAIL 受体 2（DR5）的表达来诱导不同癌细胞系的凋亡。本研究探讨了用白细胞介素-1β（IL-1β）诱导表达 TRAIL 的 hUCMSCs，再结合低剂量埃博霉素，是否能进一步诱导乳腺癌细胞系凋亡。
MTT 法检测埃博霉素对 MDA-MB-231 和 MCF-7 细胞的细胞毒性。用 Western blot 和细胞免疫荧光双重验证观察到的结果，检测细胞中感兴趣的蛋白表达。用流式细胞术检测 Annexin V/PI 凋亡法分析埃博霉素处理的乳腺癌细胞系的凋亡率，以及与乳腺癌细胞和 hUCMSCs 共培养的效果。
用 Western blot 和免疫荧光法发现，低剂量埃博霉素（2.5-5 μM）处理的乳腺癌细胞系增加了凋亡相关受体 DR4、DR5 和裂解的半胱天冬酶 8、9 和 3 的表达。此外，白细胞介素-1β诱导的 hUCMSCs 中 TRAIL 表达增强。结合这些观察结果，我们预期将白细胞介素-1β诱导的 hUCMSCs 与低剂量埃博霉素处理的 MDA-MB-231 和 MCF-7 细胞共培养可能会增强乳腺癌细胞的凋亡。我们通过流式细胞术证实，白细胞介素-1β诱导的 TRAIL 表达 hUCMSCs 与埃博霉素处理的 MDA-MB-231 和 MCF-7 细胞共培养，可提高这些乳腺癌细胞的凋亡率。
我们发现，埃博霉素上调 DR4 和 DR5 的表达，增加了乳腺癌细胞系中 TRAIL 介导的凋亡。低剂量埃博霉素处理的乳腺癌细胞系与白细胞介素-1β诱导的 TRAIL 表达 hUCMSCs 联合使用可能成为一种潜在的抗肿瘤治疗方法。
