Molecular Oncology Laboratory, Division of Oncology, Department of Medicine, University of Patras, 26504 Patras, Greece.
Department of Biomedical Sciences, University of West Attica, 12243 Athens, Greece.
Genes (Basel). 2021 Oct 23;12(11):1686. doi: 10.3390/genes12111686.
We have previously shown that overexpression of RANK-c in ER-negative breast cancer cell lines attenuates aggressive properties of cancer cells, partially through a RANK-c/EGFR interaction. EGFR inhibition through TKIs in breast cancer has been tested in triple-negative disease settings with limited clinical benefit for patients. Here we test if expression of RANK-c in ER-negative breast cancer cells in conjunction with treatment with TK inhibitors (erlotinib or gefitinib) can affect survival and colony-forming capacity of cancer cells.
Stably expressing MDA-MB-231-RANK-c and SKBR3-RANK-c cells were employed to test proliferation and colony formation in the presence of TKIs. In addition, Western blot analysis was performed to dissect EGFR related signaling cascades upon TK inhibition in the presence of RANK-c.
Interestingly the two RANK-c expressing, ER-negative cells lines presented with a distinct phenotype concerning TKI sensitivity upon treatment. MDA-MB-231-RANK-c cells had a higher sensitivity upon gefitinib treatment, while erlotinib decreased the proliferation rate of SKBR3-RANK-c cells. Further, colony formation assays for MDA-MB-231-RANK-c cells showed a decrease in the number and size of colonies developed in the presence of erlotinib. In addition, RANK-c seems to alter signaling through EGFR after TKI treatment in a cell type-specific manner.
Our results indicate that ER-negative breast cancer cells that express RANK-c alter their sensitivity profile against tyrosine kinase inhibitors (erlotinib and gefitinib) in a cell type-specific and culture substrate-dependent manner.
我们之前已经证明，在 ER 阴性乳腺癌细胞系中过表达 RANK-c 可以减弱癌细胞的侵袭性，部分是通过 RANK-c/EGFR 相互作用。在三阴性疾病环境中，已经测试了针对 EGFR 的 TKI（酪氨酸激酶抑制剂）抑制作用，但对患者的临床获益有限。在这里，我们测试 ER 阴性乳腺癌细胞中 RANK-c 的表达与 TKI（厄洛替尼或吉非替尼）治疗相结合是否会影响癌细胞的存活和集落形成能力。
使用稳定表达 MDA-MB-231-RANK-c 和 SKBR3-RANK-c 的细胞来测试 TKI 存在下的增殖和集落形成。此外，还进行了 Western blot 分析，以剖析在 RANK-c 存在下 TKI 抑制时 EGFR 相关信号级联。
有趣的是，这两个表达 RANK-c 的 ER 阴性细胞系在 TKI 敏感性方面表现出明显不同的表型。在 gefitinib 治疗下，MDA-MB-231-RANK-c 细胞的敏感性更高，而 erlotinib 降低了 SKBR3-RANK-c 细胞的增殖率。此外，MDA-MB-231-RANK-c 细胞的集落形成测定显示，在用 erlotinib 处理时，集落的数量和大小减少。此外，RANK-c 似乎以细胞类型特异性的方式改变 TKI 治疗后的 EGFR 信号传导。
我们的结果表明，表达 RANK-c 的 ER 阴性乳腺癌细胞以细胞类型特异性和培养基质依赖性的方式改变其对酪氨酸激酶抑制剂（厄洛替尼和吉非替尼）的敏感性谱。
