Detecting plasma hsa_circ_0061276 in patients with gastric cancer by reverse transcription-digital polymerase chain reaction

Abstract

BackgroundThe role of circular RNAs (circRNAs) in the occurrence of gastric cancer is still unclear. Therefore, the diagnostic value and mechanisms underlying hsa_circ_0061276 in the occurrence of gastric cancer were explored.MethodsReverse transcription-droplet digital polymerase chain reaction was used to detect the copy number of hsa_circ_0061276 in plasma from healthy individuals, as well as from patients with gastric precancerous lesions or early-stage or advanced gastric cancer. Plasmids overexpressing or knocking down hsa_circ_0061276 expression were transfected into gastric cancer cells. The effects on the growth, migration, and cell cycle distribution of gastric cancer cells were then analyzed. Finally, miRanda and RNAhybrid were used to explore the binding sites between hsa_circ_0061276 and microRNAs (miRNAs). A double luciferase reporter gene assay was used to confirm the miRNA sponge effect.ResultsThe results show that plasma hsa_circ_0061276 copy number showed a trend of a gradual decrease when comparing healthy controls to the early cancer group and advanced gastric cancer group. Overexpression of hsa_circ_0061276 inhibited the growth and migration of gastric cancer cells. Through bioinformatic analyses combined with cellular experiments, it was found that hsa_circ_0061276 inhibited the growth of gastric cancer by binding to hsa-miR-7705.ConclusionHsa_circ_0061276 may be a new biomarker for gastric cancer. The tumor suppressor role of hsa_circ_0061276 on gastric cancer likely occurs through a sponge effect on miRNAs such as hsa-miR-7705.

Current advances in circular RNA detection and investigation methods: Are we running in circles?

“…This is particularly useful regarding normalizing circRNA expression, as there is no current consensus regarding the choice of a suitable housekeeping gene, especially in RNAse R+‐treated samples (Zhong et al, 2019). So far, ddPCR has been proven helpful in quantifying circRNAs from the plasma of patients with different oncological malignancies (T. Li et al, 2018; Ruan et al, 2022; Tao et al, 2023).…”

CircRNAs in osteoarthritis: research status and prospect

“…CircRNAs are less susceptible to degradation by RNase R due to their lack of 3′and 5′ends, which confers a longer half-life and stability than other noncoding RNAs or mRNAs, suggesting the possibility of circRNAs as indicators for clinical testing. CircRNA as a liquid biopsy indicator has been frequently mentioned in tumor studies ( Li et al, 2022a ; Wang et al, 2022a ; Ruan et al, 2022 ; Xue et al, 2022 ). CircRNA was detected in cartilage, synovial fluid, and serum in OA patients and was different from the control group, suggesting that it is an indicator of early diagnosis of osteoarthritis.…”