Long noncoding RNA BBOX1-AS1 increased radiotherapy sensitivity in colorectal cancer by stabilizing and activating PFK1

Qi Wang; Xiao-Fei Li; Ying-Hui Zhou; Xiang-Hong Qin; Li-Hui Wang; Meng-Qing Xiao; Ke Cao; John K Ma; Cheng-Hui Huang

doi:10.1016/j.tranon.2023.101751

Long noncoding RNA BBOX1-AS1 increased radiotherapy sensitivity in colorectal cancer by stabilizing and activating PFK1

Transl Oncol. 2023 Oct:36:101751. doi: 10.1016/j.tranon.2023.101751. Epub 2023 Aug 4.

Authors

Qi Wang¹, Xiao-Fei Li², Ying-Hui Zhou¹, Xiang-Hong Qin¹, Li-Hui Wang¹, Meng-Qing Xiao¹, Ke Cao¹, John K Ma³, Cheng-Hui Huang⁴

Affiliations

¹ Department of Oncology, The Third Xiangya Hospital, Central South University, Changsha 410013, Hunan, China.
² Department of Oncology, The Third Xiangya Hospital, Central South University, Changsha 410013, Hunan, China; The Second Affiliated Hospital of Chengdu Medical College (China National Nuclear Corporation 416 Hospital) , Chengdu, 610051, Sichuan, China.
³ Cotton O'Neil Cancer Center, Stormont Vail Hospital, Topeka, KS, USA.
⁴ Department of Oncology, The Third Xiangya Hospital, Central South University, Changsha 410013, Hunan, China. Electronic address: hchdoctor@sohu.com.

Abstract

Purpose: Our study explored the effect of long noncoding RNA BBOX1-AS1 on colorectal cancer (CRC) radiosensitivity in vivo and in vitro.

Methods: Differentially expressed lncRNAs in CRC were screened using a bioinformatics database and an online prediction website. The expression of BBOX1-AS1 in tissue samples was analyzed via real-time quantitative PCR (RT-qPCR). Subcellular localization of BBOX1-AS1 in CRC cells was analyzed using fluorescence in situ hybridization (FISH). The correlation between BBOX1-AS1 and PFK1 expression levels in CRC tissues was analyzed via Pearson's correlation coefficient. The effect of BBOX1-AS1 on PFK1 stability was investigated using RNA and protein stability testing. RNA Binding Protein Immunoprecipitation (RIP) and RNA pull-down assays were used to confirm the binding of BBOX1-AS1 to PFK1.

Results: BBOX1-AS1 was highly expressed in CRC and associated with poor prognosis. Similarly, it was highly expressed in CRC tissues and CRC cell lines. In addition, BBOX1-AS1 promoted the proliferation, invasion, migration, and glycolysis of CRC cells and inhibited apoptosis. RIP and RNA pull-down experiments confirmed that BBOX1-AS1 bound to PFK1. RNA stability and protein stability experiments showed that BBOX1-AS1 affected the stability of PFK1 mRNA and protein. Furthermore, we confirmed that BBOX1-AS1 increased radiation resistance through the regulation of PFK1 expression.

Conclusions: BBOX1-AS1 promoted the proliferation, invasion, migration, and glycolysis of CRC cells through stabilization of the expression of PFK1. BBOX1-AS1 also inhibited CRC cell apoptosis and increased radiotherapy resistance in CRC cells.

Keywords: BBOX1-AS1; Colo205 cells; Colorectal cancer; HCT116 cells; Phosphofructokinase-1; Radiotherapy.