High-mobility group protein B1 (HMGB1) and its potential in diagnosis and treatment of ovarian cancer
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摘要:
目的 研究高迁移率族蛋白（HMGB1）在上皮性卵巢癌、卵巢良性疾病及健康人血清中的表达情况及与临床治疗的关系，通过RNA干扰抑制探讨HMGB1对卵巢癌细胞增殖、迁移和侵袭的影响。 方法 ELISA检测47例卵巢癌患者手术前及术后1个月血清中HMGB1表达水平，30例卵巢良性疾病患者作为良性肿瘤组，30例健康女性作为正常对照组；靶向HMGB1基因的慢病毒载体转染卵巢癌细胞，并利用RT-PCR和Western Blot方法检测干扰效果，CCK-8法检测细胞增殖情况，Transwell小室模型检测细胞侵袭迁移能力。 结果 卵巢癌组HMGB1水平明显高于良性肿瘤组与正常对照组（P < 0.01），卵巢癌术后HMGB1较术前明显降低（P < 0.01），抑制HMGB1的表达可以降低卵巢癌细胞的增殖、侵袭、迁移能力。 结论 HMGB1水平与卵巢癌进展密切相关，其表达下调可显著抑制卵巢癌细胞的增殖和迁移侵袭能力，有望对卵巢癌的临床检测及治疗提供新思路。 Abstract:Objective The objective of this research is to study the serum level of the high-mobility group protein B1 (HMGB1) in human ovarian tumor (OvCa) and in a healthy control. This study also aims to identify different HMGB1 levels before and after surgery and to explore the inhibitory effect of HMGB1 gene silencing in the proliferation and invasion ability of OvCa. Methods Enzyme-linked immunosorbent assay was used to measure the serum level of HMGB1 in OvCa patients and healthy subjects. Lentivirus vector with HMGB1 shRNA was constructed and used to infect OvCa cells. The expressions of HMGB1 mRNA and protein were tested by real-time PCR and Western blot. Cell proliferation was detected using the Cell Counting Kit-8 assay, whereas cell invasion and migration were detected by Transwell assay. Results The serum level of HMGB1 was more elevated in patients with malignant diseases compared with individuals with benign diseases and the control groups. In the malignant group, the serum level of HMGB1 decreased noticeably after therapy. Down-regulation of HMGB1 expression resulted in the inhibition of the biological behavior and metastasis of ovarian cancer cells. Conclusion HMGB1 is closely associated with clinicopathologic features of OvCa. Knockdown of HMGB1 expression can significantly inhibit cell proliferation, cell migration, and cell invasion of OvCa. These findings indicate that HMGB1 can function as a therapeutic target for ovarian neoplasm in the future. -
Key words:
- high mobility group protein /
- ovarian neoplasm /
- RNA interference
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图 1 HMGB1 shRNA转染后Real-time PCR和Western Blot结果
A. RT-PCR；B. Western Blot
Figure 1. Real-time PCR and Western blot results of ovarian cancer cells transfected with HMGB1 shRNA
图 2 HMGB1 shRNA转染对卵巢癌细胞生长的影响
A. SKOV3；B. A2780
Figure 2. Effect of HMGB1 shRNA transfection on ovarian cancer cell growth
图 3 HMGB1 shRNA转染后对卵巢癌细胞迁移能力的影响
Figure 3. Effect of in vitro HMGB1 shRNA transfection on migration ability of human ovarian cancer cells
图 4 HMGB1 shRNA转染后对卵巢癌细胞侵袭能力的影响
Figure 4. Effect of in vitro HMGB1 shRNA transfection in human ovarian cancer cell invasion ability
表 1 不同临床病理特征卵巢癌患者HMGB1含量比较
Table 1. Comparison of HMGB1 content in clinical cases with different pathologic characteristics
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