Overexpression of HMGB3 promotes proliferation and metastasis in esophageal squamous cell carcinoma (ESCC) and indicates a poor prognosis (IMAGE)
Caption
(A) Quantitative PCR analysis indicates the expression of HMGB3 in 20 pairs of ESCC tissues and their corresponding paratumor tissues. (B) Quantitative PCR analysis was utilized to determine the relative mRNA expression of HMGB3 in ESCC cells. (C) Western blotting was performed to evaluate the HMGB3 and TGIF2 protein levels in ESCC cells. (D) Representative immunohistochemistry images for HMGB3 staining in ESCC and corresponding paratumor tissues. (E) Semi-quantitative analysis was utilized to assess the immunohistochemistry scores of HMGB3 in both ESCC and their corresponding paratumor tissues (n = 57 pairs). (F) Kaplan–Meier analysis demonstrates the association between the mRNA expression of HMGB3 and overall survival in ESCC patients (n = 112). The red line indicates patients with relatively higher HMGB3, whereas the green line indicates the group with lower HMGB3. (G) CCK-8 assay demonstrates the alteration in the proliferative ability of ESCC cells upon HMGB3 overexpression, with optical density (OD) measurements recorded daily for 5 days. (H) Colony formation assay illustrates the proliferative ability of ESCC cells following the overexpression of HMGB3. The left panel depicts the results, whereas the right panel illustrates the number of counted colonies within the designated groups. (I) The impact of HMGB3 overexpression on the cell cycle was evaluated by flow cytometry (left), with percentages of cells in the S, G1, and G2 phases determined and compared (right). (J) Transwell analysis demonstrates the migratory and invasive capacity of ESCC cells following the overexpression of HMGB3 (left). The numbers of migrated and invasive cells were quantified and compared (right). (K–M) Nude mice, categorized into four groups, each consisting of five mice, were injected with the designated cells. (K) Images depicting the xenograft tumors. (L, M) Tumor volume and weight data from xenograft mouse tumors. (N–P) Nude mice, categorized into four groups, each consisting of 10 mice, were administered the specified cells via subcutaneous injection through the tail vein. (N) The number of metastatic nodules in the lungs was determined for each group. (O) Overall survival of the nude mice in the indicated four groups. (P) Representative hematoxylin-eosin staining images of lung tissues from the specified groups are presented. All data were expressed as mean ± standard deviation. Statistical significance is indicated as ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.
Credit
Liaoran Niu, Wanli Yang, Wei Zhou, Lili Duan, Qi Wang, Xiaoqian Wang, Yiding Li, Chengchao Xu, Yujie Zhang, Jinqiang Liu, Jian Zhang, Daiming Fan, Jianyong Zheng, Liu Hong
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