Ma Qiming, Li Xingxing, Wang Huming, Xu Shenglin, Que Yukang, He Peng, Yang Rui, Wang Qiwei, Hu Yong
Department of Orthopedics, The First Affiliated Hospital of Anhui Medical University, Hefei, 230022, Anhui, China.
Department of Orthopedics, Lu 'an Hospital of Anhui Medical University, Lu'an, 237008, Anhui, China.
Heliyon. 2024 Apr 29;10(9):e30445. doi: 10.1016/j.heliyon.2024.e30445. eCollection 2024 May 15.
To investigate the involvement of the homeobox gene B5 (HOXB5) in the progression and metastasis of osteosarcoma.
The expression of HOXB5 in human osteosarcoma tissues and its correlation with clinical indicators were investigated using bioinformatics analysis and immunohistochemical labelling. Human osteosarcoma cells (HOS, MG63, U2OS, and Saos-2) and normal human osteoblasts (hFOB1.19) were cultivated. The expression of HOXB5 in these cells was detected using western blotting (WB) and RT‒PCR. Two cell lines exhibiting elevated HOXB5 expression were chosen and divided into three groups: the blank group (mock), control group (control) and transfection group (shHOXB5). The transfection group was infected with lentivirus expressing shRNAs targeting HOXB5. The transfection efficiency was detected by WB. Cell proliferation suppression was measured by CCK-8 and 5-ethynyl-2'-deoxyuridine (EdU) assays; the percentage of apoptotic cells was determined by flow cytometry; and cell migration and invasion were detected via the Transwell chamber test. WB was utilized to determine the protein expression of genes linked to metastasis (MMP2, MMP9), apoptosis (Bax, Bcl-2), and the JAK2/STAT3 pathway (JAK2, -JAK2, STAT3, p-STAT3).
In osteosarcoma tissues, HOXB5 expression was elevated and strongly correlated with distant metastasis. Silencing HOXB5 reduced the proliferation, migration and invasion of osteosarcoma cells; prevented the progression and metastasis of tumours in tumour-bearing nude mice; and reduced the activation of key proteins in the JAK2/STAT3 signalling pathway.
Through the JAK2/STAT3 signalling pathway, HOXB5 plays a crucial role in the malignant progression of osteosarcoma and is a promising target for osteosarcoma treatment.
探讨同源框基因B5（HOXB5）在骨肉瘤进展和转移中的作用。
采用生物信息学分析和免疫组织化学标记法，研究HOXB5在人骨肉瘤组织中的表达及其与临床指标的相关性。培养人骨肉瘤细胞（HOS、MG63、U2OS和Saos-2）和正常人成骨细胞（hFOB1.19）。采用蛋白质免疫印迹法（WB）和逆转录-聚合酶链反应（RT-PCR）检测这些细胞中HOXB5的表达。选择两个HOXB5表达升高的细胞系，分为三组：空白组（mock）、对照组（control）和转染组（shHOXB5）。转染组用表达靶向HOXB5的短发夹RNA（shRNA）的慢病毒感染。通过WB检测转染效率。采用细胞计数试剂盒-8（CCK-8）和5-乙炔基-2'-脱氧尿苷（EdU）检测法测定细胞增殖抑制率；通过流式细胞术测定凋亡细胞百分比；通过Transwell小室试验检测细胞迁移和侵袭能力。利用WB检测与转移相关基因（基质金属蛋白酶2、基质金属蛋白酶9）、凋亡相关基因（Bax、Bcl-2）以及JAK2/信号转导和转录激活因子3（STAT3）通路相关基因（JAK2、-JAK2、STAT3、磷酸化STAT3）的蛋白表达。
在骨肉瘤组织中，HOXB5表达升高，且与远处转移密切相关。沉默HOXB5可降低骨肉瘤细胞的增殖、迁移和侵袭能力；抑制荷瘤裸鼠肿瘤的进展和转移；并降低JAK2/STAT3信号通路中关键蛋白的激活。
HOXB5通过JAK2/STAT3信号通路在骨肉瘤的恶性进展中起关键作用，是骨肉瘤治疗的一个有前景的靶点。
