Han Weiwei, Niu Lin, Wang Lin, Liu Jixiang, Li Huanying
Department of Otolaryngology Head and Neck Surgery, Tianjin Union Medicine Centre, Tianjin, China.
Arch Med Sci. 2019 Dec 31;17(5):1164-1174. doi: 10.5114/aoms.2019.91352. eCollection 2021.
This study was performed to explore the function of B-Raf proto-oncogene-activated non-coding RNA (BANCR) in laryngeal squamous cell carcinoma (LSCC) and cisplatin resistance.
The relative expression level of long non-coding RNA (lncRNA) BANCR was examined by qRT-PCR in tumor tissues and adjacent tissues, normal laryngeal cells (Het-1A) and laryngeal squamous carcinoma cells (TU686, TU177). Cisplatin-resistant laryngeal squamous carcinoma cell lines (TU686-DDP-R, TU177-DDP-R) were established. Next, we inhibited BANCR expression by transfecting siRNA-BANCR and enhanced BANCR expression by transfecting pcDNA3.1-BANCR into TU686, TU177, TU686-DDP-R and TU177-DDP-R cells. The CCK-8 assay and clone formation assay were performed to detect colony proliferation ability and formation ability of cells. Further, to investigate through which BANCR cell viability/formation is regulated, we detected the expression of MRP1, Bcl-2, p-PKB, and Bax by western blot.
BANCR was highly expressed in laryngeal squamous carcinoma tissues and cells. Chemoresistance was generated in TU686-DDP-R and TU177-DDP-R compared with TU686 and TU177 cells after cisplatin treatment. In addition, upregulated lncRNA BANCR reduced or postponed cell sensitivity to cisplatin by enhancing cell proliferation in TU686 and TU177 cells. Meanwhile, the expression of MRP1, Bcl-2, and p-PKB was increased, while Bax was reduced. After cisplatin treatment, down-regulation of BANCR could consequently attenuate TU686-DDP-R and TU177-DDP-R cell proliferation, and the expression of MRP1, Bcl-2, and p-PKB was decreased and Bax was increased.
Down-regulation of BANCR reverses cisplatin resistance of cisplatin-resistant LSCC cell lines.
本研究旨在探讨B-Raf原癌基因激活的非编码RNA（BANCR）在喉鳞状细胞癌（LSCC）及顺铂耐药中的作用。
采用qRT-PCR检测肿瘤组织与癌旁组织、正常喉细胞（Het-1A）及喉鳞状癌细胞（TU686、TU177）中长链非编码RNA（lncRNA）BANCR的相对表达水平。建立顺铂耐药的喉鳞状癌细胞系（TU686-DDP-R、TU177-DDP-R）。接下来，通过转染siRNA-BANCR抑制BANCR表达，转染pcDNA3.1-BANCR增强TU686、TU177、TU686-DDP-R和TU177-DDP-R细胞中的BANCR表达。进行CCK-8检测和克隆形成检测以检测细胞的集落增殖能力和形成能力。此外，为研究BANCR通过何种方式调节细胞活力/形成，通过蛋白质免疫印迹法检测MRP1、Bcl-2、p-PKB和Bax的表达。
BANCR在喉鳞状癌组织和细胞中高表达。顺铂处理后，与TU686和TU177细胞相比，TU686-DDP-R和TU177-DDP-R产生了化学抗性。此外，lncRNA BANCR上调通过增强TU686和TU177细胞增殖降低或延迟了细胞对顺铂的敏感性。同时，MRP1、Bcl-2和p-PKB的表达增加，而Bax减少。顺铂处理后，BANCR下调可减弱TU686-DDP-R和TU177-DDP-R细胞增殖，MRP1、Bcl-2和p-PKB的表达降低，Bax增加。
BANCR下调可逆转顺铂耐药的LSCC细胞系的顺铂耐药性。
