Seong Hee Kyung, Osaki Mitsuhiko, Izutsu Runa, Jehung Jumond P, Sato Reo, Sasaki Ryo, Hamada Junichi, Yamamoto Atsushi, Takenaka Atsushi, Okada Futoshi
Division of Experimental Pathology, Faculty of Medicine, Tottori University, 86 Nishicho, Yonago, 683-8503, Japan.
Chromosome Engineering Research Center, Tottori University, Yonago, 683-8503, Japan.
Sci Rep. 2025 Sep 25;15(1):32861. doi: 10.1038/s41598-025-18095-7.
Amphoterin-induced gene and open reading frame 2 (AMIGO2) was previously identified as a driver gene for liver metastasis. AMIGO2 has also been found to be a prognostic factor for cancer patients with a low frequency of spontaneous liver metastasis. Here, we investigated whether AMIGO2 expression affects the acquisition of cancer stem cell-like properties in cancer cells by using human gastric (MKN45) and colon (DLD-1) cancer cell lines. Knockdown of AMIGO2 expression in MKN45 and DLD-1 cells via transfection with short hairpin RNA-AMIGO2 suppressed cell proliferation under several starvation conditions in two-dimensional culture. AMIGO2 knockdown cells showed reduced ability to form multicellular spheres in a three-dimensional culture. Half-maximal inhibitory concentration values of anticancer drugs in sphere-forming cells were decreased by AMIGO2 knockdown. Silencing of AMIGO2 suppressed tumor formation and growth of subcutaneously injected tumors in NOD/SCID mice. Expression levels of AMIGO2 and cancer stem cell markers, such as CD44, CD133, and EpCAM, were almost identical in spherically grown cancer cells. AMIGO2 expression was positively correlated with expression of existing cancer stem cell markers in multiple organ cancer cell lines. These results demonstrate that AMIGO2 accelerates the malignant progression of human cancer cells by inducing a cancer stem cell-like phenotype.
双调蛋白诱导基因及开放阅读框2（AMIGO2）先前被鉴定为肝转移的驱动基因。AMIGO2也被发现是自发肝转移发生率低的癌症患者的一个预后因素。在此，我们通过使用人胃癌（MKN45）和结肠癌细胞系（DLD-1），研究了AMIGO2表达是否影响癌细胞中癌症干细胞样特性的获得。通过用短发夹RNA-AMIGO2转染敲低MKN45和DLD-1细胞中的AMIGO2表达，在二维培养的几种饥饿条件下抑制了细胞增殖。AMIGO2敲低的细胞在三维培养中形成多细胞球的能力降低。AMIGO2敲低降低了成球细胞中抗癌药物的半数最大抑制浓度值。AMIGO2的沉默抑制了NOD/SCID小鼠皮下注射肿瘤的形成和生长。在球形生长的癌细胞中，AMIGO2与癌症干细胞标志物如CD44、CD133和EpCAM的表达水平几乎相同。在多个器官癌细胞系中，AMIGO2表达与现有癌症干细胞标志物的表达呈正相关。这些结果表明，AMIGO2通过诱导癌症干细胞样表型加速人癌细胞的恶性进展。
