DataSheet_1_LncRNA KASRT Serves as a Potential Treatment Target by Regulating SRSF1-Related KLF6 Alternative Splicing and the P21/CCND1 Pathway in Osteosarcoma: An In Vitro and In Vivo Study.zip

PurposeLong non-coding RNA KLF6 alternative splicing regulating transcript (lnc-KASRT) locates within the intronic region of SRSF1, possessing the potential to regulate KLF6 alternative splicing to promote carcinogenicity. Then, the current in vitro and in vivo study aimed to investigate the effect of lnc-KASRT on regulating tumor malignant behaviors, and the implication of its interaction with KLF6 alternative splicing in osteosarcoma.MethodsLnc-KASRT overexpression or knockdown plasmid was transfected into U-2OS and Saos-2 cells. Then, KLF6-SV1 knockdown plasmid with or without lnc-KASRT overexpression plasmid was transfected into these cells for compensative experiments. In vivo, lnc-KASRT overexpression or knockdown Saos-2 cells were injected in mice for tumor xenograft construction.ResultsLnc-KASRT expression was increased in most osteosarcoma cell lines compared to control cell line. Lnc-KASRT overexpression promoted cell viability, mobility, and anti-apoptotic marker expression, while reducing apoptosis rate and pro-apoptotic marker expression; meanwhile, it regulated SRSF1, KLF6 alternative splicing (increased KLF6-splice variant 1 (KLF6-SV1), decreased KLF6-wild type (KLF6-WT)), and followed P21/CCND1 pathway in U-2OS/Saos-2 cells. The lnc-KASRT knockdown exhibited opposite trends. Subsequent compensative experiments disclosed that KLF6-SV1 knockdown attenuated most of the tumor-promoting effects of lnc-KASRT overexpression in U-2OS/Saos-2 cells. In vivo experiments further validated that lnc-KASRT enhanced tumor growth and reduced tumor apoptosis; meanwhile, it also increased tumor KLF6-SV1, MMP-1, and MMP-9 expressions but decreased tumor SRSF1 and KLF6-WT expressions in xenograft mice.ConclusionLnc-KASRT serves as a potential treatment target via regulating SRSF1-related KLF6 alternative splicing and following P21/CCND1 pathway in osteosarcoma.

旨在研究长非编码RNA KLF6可变剪接调控转录物（lnc-KASRT）位于SRSF1的内含子区域，具有调节KLF6可变剪接以促进致癌性的潜力。随后，本研究旨在通过体外和体内实验探究lnc-KASRT对调节肿瘤恶性表型的影响，以及其在骨肉瘤中与KLF6可变剪接相互作用的意义。
方法：将lnc-KASRT过表达或敲低载体转染至U-2OS和Saos-2细胞。随后，为进行补偿实验，将KLF6-SV1敲低载体与lnc-KASRT过表达载体联合转染至这些细胞。体内实验中，将lnc-KASRT过表达或敲低Saos-2细胞注射至小鼠体内构建肿瘤异种移植。
结果：与对照细胞系相比，大多数骨肉瘤细胞系中lnc-KASRT的表达增加。lnc-KASRT过表达促进了细胞活力、迁移能力和抗凋亡标记物的表达，同时降低了凋亡率和促凋亡标记物的表达；同时，它在U-2OS/Saos-2细胞中调节了SRSF1、KLF6可变剪接（KLF6-splice variant 1 (KLF6-SV1) 的增加，KLF6-wild type (KLF6-WT) 的减少），并遵循P21/CCND1通路。lnc-KASRT敲低呈现相反趋势。后续的补偿实验发现，KLF6-SV1敲低减弱了lnc-KASRT过表达在U-2OS/Saos-2细胞中的大多数促肿瘤效应。体内实验进一步证实，lnc-KASRT增强了肿瘤生长并减少了肿瘤凋亡；同时，它还增加了异种移植小鼠肿瘤中KLF6-SV1、MMP-1和MMP-9的表达，但降低了SRSF1和KLF6-WT的表达。
结论：通过调节与SRSF1相关的KLF6可变剪接并遵循P21/CCND1通路，lnc-KASRT成为骨肉瘤治疗的一个潜在靶点。