Abstract
Objective: Emerging evidence indicates that long non-coding RNA (lncRNA) RP11-93B14.5 facilitates tumor progression in variety of malignancies. The present study proposed to study the functional effect of lncRNA RP11-93B14.5 in gastric cancer (GC) as well as the underlying mechanism. Methods: Bioinformatics analysis was utilized to analyze lncRNA expression in GC tissues. siRNA was used for knockdown of RP11-93B14.5 in GC cells MKN45 and KATO III. The stable knockdown cell lines were constructed by CRISPR-Cas9. Cell counting kit-8 (CCK-8) assay and soft agar colony formation assay were used to analyze GC cell viability. Flow cytometry analysis was performed to analyze the cell cycle distribution of MKN45 and KATO III. RNA sequencing (RNA-seq) was employed to detect differential genes after transfection with siRP11-93B14.5. Quantitative PCR (Q-PCR) was used to examine gene expression in GC cell lines. Western-blot assay was used to measure protein levels. RNA fluorescent in situ hybridization (FISH) was conducted for lncRNA cellular location and expression. Results: Based on the Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) database, RP11-93B14.5 was upregulated in GC tissue, which was also verified in GC cell lines in comparison to the normal gastric epithelial HFE145 cells. Knockdown of RP11-93B14.5 decreased cell viability and the colony number of MKN45 and KATO III cells, and altered cell cycle distribution in vitro. RNA-seq analysis revealed RP11-93B14.5 may modulate genes expression of S100A2 and TIMP2 in MKN45 and KATO III cells. Mechanistically, RP11-93B14.5 may drive the progression of GC via S100A2 related-PI3K/AKT signaling pathway. Conclusions: LncRNA RP11-93B14.5 knockdown alleviated the malignant phenotypes of GC cells through regulating PI3K/AKT. Our results provide evidence for the role of lncRNAs in regulating tumor progression.
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The data used to support the findings of the present study are available from the corresponding author upon reasonable request.
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Funding
This work was supported by the National Natural Science Foundation of China (No. 81803237), the grant from Natural Science Foundation of Sichuan Province, China (No. 2023NSFSC1848), the grant from Natural Science Foundation of Anhui Provincial Education Department (No. KJ2019A0421) and the grant from Science and Technology Program of Luzhou, China (No. 2022-JYJ-113).
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ML and YJ designed and conceived the study. XW, FD, WL, FW, LG and YS conducted the bioinformatics analysis. XW, QL, ZZ, HZ, HY, LL, MC, YZ, YC, JS and ZX performed the experiments. XW, QL, HY, XL, YZ, WL, YC, JS, ZX and LG analyzed the data. YC, JS, ZX, YS and MC provided the resources of experiments for this study. QL wrote the original draft. ML, XW, ZZ and LL reviewed and edited the manuscript. All authors read and approved the final manuscript.
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Supplementary Figure S1:
CRISPR-Cas9 induced knockdown of lncRNA RP11-93B14.5 suppressed GC cell proliferation through modulating PI3K/AKT pathway.
Supplementary Table S1:
Primer sequences and target sequences used in the present study.
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Li, Q., Zhu, Z., Zhang, H. et al. LncRNA RP11-93B14.5 promotes gastric cancer cell growth through PI3K/AKT signaling pathway. Mol Biotechnol (2023). https://doi.org/10.1007/s12033-023-00844-6
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DOI: https://doi.org/10.1007/s12033-023-00844-6