A novel circular RNA hsa_circRNA_103809/miR-377-3p/GOT1 pathway regulates cisplatin-resistance in non-small cell lung cancer (NSCLC)

Abstract

Background Cisplatin is the first-line chemotherapeutic drug for non-small cell lung cancer (NSCLC), and emerging evidences suggests that targeting circular RNAs (circRNAs) is an effective strategy to increase cisplatin-sensitivity in NSCLC, but the detailed mechanisms are still not fully delineated. Methods Cell proliferation, viability and apoptosis were examined by using the cell counting kit-8 (CCK-8) assay, trypan blue staining assay and Annexin V-FITC/PI double staining assay, respectively. The expression levels of cancer associated genes were measured by using the Real-Time qPCR and Western Blot analysis at transcriptional and translated levels. Dual-luciferase reporter gene system assay was conducted to validated the targeting sites among hsa_circRNA_103809, miR-377-3p and 3′ untranslated region (3’UTR) of GOT1 mRNA. The expression status, including expression levels and localization, were determined by immunohistochemistry (IHC) assay in mice tumor tissues. Results Here we identified a novel hsa_circRNA_103809/miR-377-3p/GOT1 signaling cascade which contributes to cisplatin-resistance in NSCLC in vitro and in vivo. Mechanistically, parental cisplatin-sensitive NSCLC (CS-NSCLC) cells were subjected to continuous low-dose cisplatin treatment to generate cisplatin-resistant NSCLC (CR-NSCLC) cells, and we found that hsa_circRNA_103809 and GOT1 were upregulated, while miR-377-3p was downregulated in CR-NSCLC cells but not in CS-NSCLC cells. In addition, hsa_circRNA_103809 sponged miR-337-3p to upregulate GOT1 in CS-NSCLC cells, and knock-down of hsa_circRNA_103809 enhanced the inhibiting effects of cisplatin on cell proliferation and viability, and induced cell apoptosis in CR-NSCLC cells, which were reversed by downregulating miR-377-3p and overexpressing GOT1. Consistently, overexpression of hsa_circRNA_103809 increased cisplatin-resistance in CS-NSCLC cells by regulating the miR-377-3p/GOT1 axis. Finally, silencing of hsa_circRNA_103809 aggravated the inhibiting effects of cisplatin treatment on NSCLC cell growth in vivo. Conclusions Analysis of data suggested that targeting the hsa_circRNA_103809/miR-377-3p/GOT1 pathway increased susceptibility of CR-NSCLC cells to cisplatin, and this study provided novel targets to improve the therapeutic efficacy of cisplatin for NSCLC treatment in clinic.

Knock-down of microRNA miR-556-5p increases cisplatin-sensitivity in non-small cell lung cancer (NSCLC) via activating NLR family pyrin domain containing 3 (NLRP3)-mediated pyroptotic cell death

“…Cisplatin-resistance is a huge obstacle to make this drug ineffective to treat NSCLC in clinic [13][14][15], and uncovering the potential underlying mechanisms will help to solve this problem. According to recent publications [16,44,46,47], miRNAs with post-transcriptional activities are crucial for regulating cancer progression and drug resistance, and multiple miRNAs that regulate cisplatin-resistance in NSCLC have been screened out, such as let-7 miRNA [16], miR-497-5p [46], miR-130a [47] and miR-377-3p [44]. Among all the miRNAs, miR-556-5p has also been reported to be closely associated with cancer progression, and its role in regulating cancer development varies according to differential cancer types [23][24][25].…”

“…The parental cisplatin-sensitive NSCLC (CS-NSCLC) cells (A549 and H1299) were purchased from The Chinese Academy of Sciences Cell Bank (Shanghai, China), and the cells were cultured in the DMEM medium (Gibco, CA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, CA, USA), 1% penicillin and streptomycin (Sigma-Aldrich, St. Louis, MO, USA). The cells were placed in an incubator with 5% CO 2 humidified air at 37°C, and the CS-NSCLC cells were subjected to long-term low-dose cisplatin exposure (0.5–5 μg/ml, 80 days) to establish cisplatin-resistant NSCLC (CR-NSCLC) cells (A549/DDP, H1299/DDP) following the experimental procedures provided by the previous work [ 44 ], and the CR-NSCLC cells were maintained in the complete medium containing 1 μg/ml cisplatin, which were subsequently subjected to high-dose cisplatin (25 μg/ml) treatment and 1 mM of pyroptosis inhibitor Necrosulfonamide (NSA, HY-100573, MedChem Express) for 0 h, 6 h, 12 h, 18 h, 24 h and 48 h.…”

The Role of Circular RNAs in the Drug Resistance of Cancers

“…Accumulating evidence has demonstrated that circRNAs are implicated in CDDP resistance in lung cancer. Knockdown or silencing of certain circRNAs suppresses CDDP resistance in NSCLC such as circ-RNF121, circ_0076305, circRNA_103615, circ_0008928, hsa_circRNA_103809, circ-PRMT5, and circ_0007385 (17,18,49,(62)(63)(64)(65). Additionally, circAKT3 inhibits glycolysis balance and enhances CDDP resistance, whereas circRNA-FOXO3 accelerates glycolysis and promotes CDDP sensitivity (50,51).…”

Inorganic–inorganic nanohybrids for drug delivery, imaging and photo-therapy: recent developments and future scope

“… 70 Typically, a few inorganic-based therapeutic agents are briefly reviewed from a chemical point of view. Platinum-based therapeutic agents, such as cisplatin, 72 carboplatin 73–75 and oxaliplatin, 76–78 and a ruthenium( iii ) complex 79 have been well explored in the drug-delivery field.…”