| 梁婉琪,赵宗霞,李芬霞,常丽花.HMGB1通过上调E2F3促进卵巢癌细胞增殖和侵袭能力[J].,2020,(4):654-660 |
| HMGB1通过上调E2F3促进卵巢癌细胞增殖和侵袭能力 |
| HMGB1 Promotes Proliferation and Invasion of Ovarian Cancer Cells by Up-regulating E2F3 |
| 投稿时间：2019-06-08 修订日期：2019-06-30 |
| DOI：10.13241/j.cnki.pmb.2020.04.011 |
| 中文关键词: HMGB1 E2F3 卵巢癌 增殖 侵袭 |
| 英文关键词: HMGB1 E2F3 Ovarian cancer Proliferation Metastasis |
| 基金项目:陕西省科技统筹基金项目(2016KTZDSF01-013-04) |
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| 中文摘要: |
| 摘要 目的：探讨高迁移率族蛋白B1(High-mobility Group Box 1 protein，HMGB1)是否通过激活E2F转录调节因子3（E2F transcription factor 3，E2F3）促进卵巢癌细胞的增殖和侵袭作用。方法：选取本院2017年10月-2018年7月收治的38例卵巢癌患者。qRT-PCR、Western blot和免疫组化分别检测患者肿瘤组织和癌旁组织中HMGB1的表达；qRT-PCR和Western blot检测HMGB1在卵巢癌细胞系A2780、HO8910、ES-2和SKOV3中的表达；RNA沉默技术敲除HMGB1在SKOV3中的表达，同时CCK-8和Trans-well观察SKOV3细胞增殖和侵袭能力变化；RNA转录组测序（RNA transcriptome sequencing，RNA-seq）检测敲除HMGB1前后SKOV3细胞中mRNA的变化； qRT-PCR和Western blot检测验证RNA-seq结果；RNA沉默技术敲除E2F3在SKOV3细胞中的表达，同时CCK-8和Trans-well观察SKOV3细胞增殖和侵袭能力变化。结果：卵巢癌肿瘤组织中HMGB1表达显著高于癌旁组织(P<0.05)；HMGB1在四种卵巢癌细胞系中的SKOV3细胞中表达最高 (P<0.05)；si-RNA基因沉默后，HMGB1在SKOV3细胞中几乎无表达，CCK-8和Trans-well实验表明HMGB1 si-RNA沉默后其增殖和侵袭能力显著被抑制(P<0.05)；RNA-seq检测结果表明，HMGB1 si-RNA沉默后 E2F3、叉状头转录因子3（Forkhead box protein P3，FOXP3）和趋化因子5（chemokine 5，CXCL5）下调倍数最明显其中E2F3下调倍数最高，同时在细胞系中验证结果显示同样结果(P<0.05)；si-RNA基因沉默E2F3后，E2F3在SKOV3细胞中几乎无表达，CCK-8和Trans-well实验表明E2F3 si-RNA沉默后其增殖和侵袭能力显著被抑制(P<0.05)。结论：HMGB1可能通过上调E2F3的表达增加促进卵巢癌SKOV3细胞增殖和转移。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate whether high-mobility group box protein B1 (HMGB1) promotes the proliferation and invasion of ovarian cancer cells by activating nuclear transcription factor E2F3. Methods: A total of 138 patients with ovarian cancer admitted to our hospital from June 2016 to May 1818 were enrolled. qRT-PCR, Western blot and immunohistochemistry were used to detect the expression of HMGB1 in tumor tissues and paracancer tissues. The expression of HMGB1 in ovarian cancer cell lines A2780, HO8910, ES-2 and SKOV3 was detected by qRT-PCR and Western blot. RNA silencing technique knocked out the expression of HMGB1 in SKOV3, while CCK-8 and Trans-well observed changes in SKOV3 cell proliferation and invasion ability. RNA transcriptome sequencing(RNA-seq) detected mRNA changes in SKOV3 cells before and after HMGB1 knockdown.The results of RNA-seq were verified by qRT-PCR and Western blot. The expression of E2F3 in SKOV3 cells was knocked out by RNA silencing technique. The proliferation and invasion ability of SKOV3 cells were observed by CCK-8 and Trans-well. Results: The expression of HMGB1 in ovarian cancer tissues was significantly higher than that in normal tissues(P<0.05). HMGB1 was the highest in SKOV3 cells of four ovarian cancer cell lines(P<0.05). After silencing of si-RNA gene, HMGB1 was almost no expression in SKOV3 cells. CCK-8 and Trans-well experiments showed that the proliferation and invasion ability of HMGB1 si-RNA was significantly inhibited after silencing(P<0.05). The results of RNA-seq assay showed that HMGB1 si-RNA was silenced. The down-regulation multiples of E2F3, FOXP3 and CXCL5 were the most obvious, and the E2F3 down-regulation was the highest, and the results in the cell line showed the same results(P<0.05). After silencing E2F3 by si-RNA gene, E2F3 showed almost no expression in SKOV3 cells, CCK- 8 and Trans-well experiments showed that the proliferation and invasion ability of E2F3 si-RNA was significantly inhibited(P<0.05). Conclusion: HMGB1 may promote the proliferation and metastasis of ovarian cancer SKOV3 cells by up-regulating the expression of E2F3. |
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