环状BACH1/微小RNA-7a-5p轴通过上调CREB5表达增强结直肠癌的增殖和转移。
CircBACH1/let-7a-5p axis enhances the proliferation and metastasis of colorectal cancer by upregulating CREB5 expression.
作者信息
Li Jutang, Tang Qian, Dong Wei, Wang Yizhou
机构信息
Key Laboratory of Signaling Regulation and Targeting Therapy of Liver Cancer, the Second Military Medical University, Shanghai, China.
Department of Gastroenterology, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
出版信息
J Gastrointest Oncol. 2020 Dec;11(6):1186-1199. doi: 10.21037/jgo-20-498.
BACKGROUND
In this study, we investigated the influences of circBACH1 on the proliferation, metastasis, migration, and apoptosis of human colorectal cancer LoVo cells and explored the molecular mechanism of its effect to guide the clinical diagnosis, treatment, and follow-up of colorectal cancer.
METHODS
The expression of circBACH1 in colorectal cancer cells was measured to determine the high expression of BACH1 in colorectal cancer (CRC). LoVo was selected for a follow-up experiment. Then, quantificational reverse transcription-polymerase chain reaction (qRT-PCR) and biotinylated let-7a-5p probes were used to confirm that the expression of let-7a-5p was lowered in colorectal cancer, and let-7a-5p was the downstream target of BACH1 in CRC. Cell counting Kit-8 (CCK-8), Transwell, and wound repair experiments confirmed that BACH1 augmented the proliferation, migration, and metastasis of CRC by regulating let-7a-5p. The apoptosis rate was measured by flow cytometry. It was concluded that BACH1 inhibited apoptosis by regulating let-7a-5p in CRC. The results of the bioinformatics analysis showed that CREB5 was overexpressed in CRC by qRT-PCR and Western blot. The results of qRT-PCR, CCK-8 assay, Transwell assay, and flow cytometry showed that let-7a-5p inhibited the proliferation, migration, and invasion of CRC cells through targeting CREB5 and augmented cell apoptosis. According to tumor growth and the determination of CREB5 by Western blot, BACH1 can affect the proliferation of CRC cells through CREB5.
RESULTS
Overall, our study confirmed that BACH1 and CREB5 increased, while the expression of let-7a-5p was lowered in colorectal cancer cells. These different expressions enhance the proliferation, metastasis, and migration of colorectal cancer cells and inhibit colorectal cancer cells' apoptosis.
CONCLUSIONS
Our study clearly illustrates the molecular mechanism of circBACH1 acting on colorectal cancer, which can be used as a therapeutic target to augment colorectal cancer treatment.
背景
在本研究中，我们调查了环状BACH1（circBACH1）对人结肠癌细胞LoVo增殖、转移、迁移和凋亡的影响，并探讨了其作用的分子机制，以指导结肠癌的临床诊断、治疗及随访。
方法
检测结肠癌细胞中circBACH1的表达，以确定其在结肠癌（CRC）中的高表达。选择LoVo进行后续实验。然后，采用定量逆转录-聚合酶链反应（qRT-PCR）和生物素化的let-7a-5p探针，证实let-7a-5p在结肠癌中的表达降低，且let-7a-5p是CRC中BACH1的下游靶点。细胞计数试剂盒-8（CCK-8）、Transwell和伤口修复实验证实，BACH1通过调节let-7a-5p增强CRC的增殖、迁移和转移。通过流式细胞术测量凋亡率。得出结论：BACH1在CRC中通过调节let-7a-5p抑制凋亡。生物信息学分析结果显示，通过qRT-PCR和蛋白质免疫印迹法（Western blot）证实CREB5在CRC中过表达。qRT-PCR、CCK-8检测、Transwell检测和流式细胞术结果显示，let-7a-5p通过靶向CREB5抑制CRC细胞的增殖、迁移和侵袭，并增加细胞凋亡。根据肿瘤生长情况及Western blot检测CREB5，BACH1可通过CREB5影响CRC细胞的增殖。
结果
总体而言，我们的研究证实，在结肠癌细胞中BACH1和CREB5升高，而let-7a-5p的表达降低。这些不同的表达增强了结肠癌细胞的增殖、转移和迁移，并抑制结肠癌细胞的凋亡。
结论
我们的研究清楚地阐明了circBACH1作用于结肠癌的分子机制，其可作为增强结肠癌治疗的治疗靶点。
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