Department of Oncology, Affiliated Hospital of Xiangnan University, Chenzhou, 423000, Hunan, China.
Department of Oncology, The First Affiliated Hospital of Jinan University, Guangzhou, 510630, Guangdong, China.
Cell Biochem Biophys. 2021 Jun;79(2):397-405. doi: 10.1007/s12013-021-00975-0. Epub 2021 Mar 20.
B7 homolog 3 (B7-H3), a member of the immunoregulatory ligand B7 family, is pivotal in T-cell-mediated immune response. It is widely expressed in diverse human tumors and its high expression indicates the poor prognosis of the patients. Nonetheless, B7-H3's role in colorectal cancer (CRC) needs to be further explored.
Western blot and immunohistochemistry were employed for detecting B7-H3 protein expression in CRC tissues and cell lines, respectively. Quantitative real-time polymerase chain reaction (qRT-PCR) was utilized for detecting B7-H3 mRNA and miR-128 expression levels. CRC cell lines SW620 and HT29 were used to construct B7-H3 overexpression or knockdown cell models, respectively. Cell counting kit-8 (CCK-8), 5-bromo-2'-deoxyuridine (BrdU), and scratch wound healing assays were employed for evaluating the effects of B7-H3 on CRC cell multiplication and migration. Besides, the regulatory relationship between miR-128 and B7-H3 was validated through dual-luciferase reporter gene assay, qRT-PCR, and western blotting.
B7-H3 expression level was remarkably elevated in CRC tissues and cell lines, and its high expression level was associated with increased tumor size, positive lymph node metastasis, and increased T stage. In CRC cells, B7-H3 overexpression significantly facilitated the cell multiplication and migration, while B7-H3 knockdown worked oppositely. Moreover, B7-H3 was identified as a target of miR-128, and miR-128 negatively regulated B7-H3 expression in CRC cells.
B7-H3 expression is upregulated in CRC tissues and cell lines, and B7-H3 participates in promoting the proliferation and migration of CRC cells. Besides, B7-H3 expression is negatively regulated by miR-128 in CRC.
B7 同源物 3（B7-H3）是免疫调节配体 B7 家族的成员，在 T 细胞介导的免疫反应中起着关键作用。它广泛表达于多种人类肿瘤中，其高表达表明患者预后不良。然而，B7-H3 在结直肠癌（CRC）中的作用仍需进一步探讨。
采用 Western blot 和免疫组织化学法分别检测 CRC 组织和细胞系中 B7-H3 蛋白的表达，采用实时定量聚合酶链反应（qRT-PCR）检测 B7-H3mRNA 和 miR-128 的表达水平。分别构建 CRC 细胞系 SW620 和 HT29 的 B7-H3 过表达或敲低细胞模型。采用细胞计数试剂盒-8（CCK-8）、5-溴-2'-脱氧尿苷（BrdU）和划痕愈合实验评估 B7-H3 对 CRC 细胞增殖和迁移的影响。此外，通过双荧光素酶报告基因检测、qRT-PCR 和 Western blot 验证 miR-128 与 B7-H3 之间的调控关系。
B7-H3 的表达水平在 CRC 组织和细胞系中显著升高，其高表达水平与肿瘤大小增加、阳性淋巴结转移和 T 分期升高有关。在 CRC 细胞中，B7-H3 过表达显著促进细胞增殖和迁移，而 B7-H3 敲低则相反。此外，B7-H3 被鉴定为 miR-128 的靶基因，miR-128 负调控 CRC 细胞中 B7-H3 的表达。
B7-H3 在 CRC 组织和细胞系中表达上调，B7-H3 参与促进 CRC 细胞的增殖和迁移。此外，miR-128 负调控 CRC 中 B7-H3 的表达。
