Zhang Shuai, Xu Siguang, Li Dandan, Wu Songxin, Han Miaomiao, Han Yifei, Wang Zixi, Qiao Dan, Yuan Hang, Du Baoshun, Chen Hongwei, Zhang Zheying
Department of Pathology, School of Basic Medical Sciences, Xinxiang Medical University Xinxiang 453003, Henan, China.
Department of Emergency Center Emergency Critical Care, The Fourth Clinical College of Xinxiang Medical College Xinxiang 453003, Henan, China.
Am J Cancer Res. 2024 Nov 15;14(11):5504-5520. doi: 10.62347/PNKH7683. eCollection 2024.
Long non-coding RNA (lncRNA)-encoded small proteins play a major role in colorectal cancer. To identify more functional encoded small proteins, ribosome profiling data from colorectal cancer (CRC) cells were screened for ribosome-associated lncRNAs. The search identified LINC01547 that was shown to encode a small protein of 76 amino acids, termed LINC01547-ORF. Real-time quantitative fluorescence showed that LINC01547 expression was downregulated in colorectal cancer tissues. However, cell functional assays revealed that LINC01547 inhibited the proliferation and migration of colorectal cancer cell lines. Meanwhile, western blot and immunofluorescence assays confirmed that LINC01547 encoded LINC01547-ORF. Cellular functional assays indicated that compared with LINC01547 itself, LINC01547-ORF inhibited the proliferation and migration of colorectal cancer cell lines. Gene set enrichment analysis identified enrichment in the focal adhesion pathway and association with CLDN18 protein, whereas protein molecular docking models revealed interacting domains and amino acid residue sites. Of note, co-immunoprecipitation and immunofluorescence experiments showed that LINC01547-ORF could bind to the CLDN18 protein and that this interaction reduced CLDN18 ubiquitination, thereby promoting protein expression. Finally, western blot and immunofluorescence assays confirmed that LINC01547-ORF could target CLDN18 to inhibit the FAK/PI3K/AKT signaling pathway, suppressing colorectal cancer cell development. These findings suggest that the LINC01547-ORF-encoded small protein inhibits proliferation and migration in colorectal cancer, thereby offering a promising direction for treating this disease.
长链非编码RNA（lncRNA）编码的小蛋白在结直肠癌中起主要作用。为了鉴定更多具有功能的编码小蛋白，对来自结直肠癌细胞的核糖体分析数据进行筛选，以寻找与核糖体相关的lncRNA。该筛选鉴定出LINC01547，它被证明可编码一种由76个氨基酸组成的小蛋白，称为LINC01547-ORF。实时定量荧光显示，LINC01547在结直肠癌组织中的表达下调。然而，细胞功能实验表明，LINC01547抑制了结直肠癌细胞系的增殖和迁移。同时，蛋白质印迹和免疫荧光实验证实LINC01547编码LINC01547-ORF。细胞功能实验表明，与LINC01547本身相比，LINC01547-ORF抑制了结直肠癌细胞系的增殖和迁移。基因集富集分析确定了粘着斑通路中的富集以及与CLDN18蛋白的关联，而蛋白质分子对接模型揭示了相互作用结构域和氨基酸残基位点。值得注意的是，免疫共沉淀和免疫荧光实验表明，LINC01547-ORF可与CLDN18蛋白结合，这种相互作用减少了CLDN18的泛素化，从而促进了蛋白表达。最后，蛋白质印迹和免疫荧光实验证实，LINC01547-ORF可靶向CLDN18以抑制FAK/PI3K/AKT信号通路，从而抑制结直肠癌细胞的发展。这些发现表明，LINC01547-ORF编码的小蛋白可抑制结直肠癌的增殖和迁移，从而为治疗这种疾病提供了一个有前景的方向。
