Overexpressed methyltransferase-like 1 (METTL1) increased chemosensitivity of colon cancer cells to cisplatin by regulating miR-149-3p/S100A4/p53 axis

Abstract

Methyltransferase-like 1 (METTL1) mediated 7-methylguanosine (m7G) is crucial for the regulation of chemoresistance in cancer treatment. However, the role of METTL1 in regulating chemoresistance of colon cancer (CC) cells to cisplatin is still unclear. This study established the cisplatin-resistant CC (CR-CC) cells and found that METTL1 was low-expressed in CR-CC cells compared to their paired cisplatin-sensitive CC (CS-CC) cells. Besides, overexpressed METTL1 enhanced the cytotoxic effects of cisplatin on CR-CC cells. In addition, miR-149-3p was the downstream target of METTL1, which could be positively regulated by METTL1. Further results validated that miR-149-3p was low-expressed in CR-CC cells comparing to the CS-CC cells. In addition, the promoting effects of overexpressed METTL1 on cisplatin induced CR-CC cell death were abrogated by synergistically knocking down miR-149-3p. Furthermore, S100A4/p53 axis was the downstream target of METTL1 and miR-149-3p, and either overexpressed METTL1 or miR-149-3p increased p53 protein levels in CR-CC cells, which were reversed by upregulating S100A4. Similarly, the promoting effects of overexpressed METTL1 on cisplatin-induced CR-CC cell death were abrogated by overexpressing S100A4. Taken together, overexpression of METTL1 sensitized CR-CC cells to cisplatin by modulating miR-149-3p/S100A4/p53 axis.

A Novel Circular RNA hsa_circRNA_103809/miR-377-3p/GOT1 Pathway Regulates Cisplatin-Resistance in Non-Small Cell Lung Cancer (NSCLC)

“…Therefore, in the present study, we identi ed a novel circRNA, hsa_circRNA_103809, that regulated cisplatin-resistance in NSCLC. Mechanistically, according to the previous publications [35,36], the cisplatin-resistant NSCLC (CR-NSCLC) cells were inducted from their corresponding parental cisplatin-sensitive NSCLC (CS-NSCLC) cells, and we found that hsa_circRNA_103809 tended to be high-expressed in CR-NSCLC cells, compared to CS-NSCLC cells. Interestingly, further experiments evidenced that knock-down of hsa_circRNA_103809 enhanced the inhibiting effects of cisplatin on cell proliferation and viability in CR-NSCLC cells.…”

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

“…The Roswell Park Memorial Institute 1640 medium (RPMI-1640, HyClone, USA) containing 10% fetal bovine serum (FBS, Gibco, USA). According to the experimental procedures provided by the previous work [35,36] and our preliminary experiments (data not shown), the CS-NSCLC cells were exposed to continuous low-dose cisplatin stimulation, ranged from 0.5 μg/ml to 5 μg/ml, for 80 days in a step-wise manner to generate descendent CR-NSCLC cells (A549/DDP, H1299/DDP and Calu-3/DDP). After that, the CR-NSCLC cells were stimulated with high-dose cisplatin (25 μg/ml) for 0 h, 24 h, 48 h and 72 h, to validate the successful induction of CR-NSCLC cells.…”

“…The NSCLC cells were subjected to differential treatments, and the TRIzol reagent (Invitrogen, USA) was employed to extract the total RNA. Next, the Real-Time qPCR was conducted to determine the expression levels of hsa_circRNA_103809, miR-377-3p and GOT1 mRNA, and the experimental procedures had all been documented in the previous publications [35,36]. Of note, to detect hsa_circRNA_103809 levels, the total RNA must be pre-treated with RNase R enzyme (3 U/μg) for 20 min at 37 ℃ to eliminate linear RNA.…”

“…The RIPA lysis buffer was purchased from Solarbio (Beijing, China) to lyse the NSCLC cells/tissues and extract the total protein, according to the experimental procedures recorded in the previous publications [35,36], the expression levels of GOT1, β-actin, cyclin D1, CDK2, cleaved caspase-3 and Bax were examined by using the Western Blot analysis. The 40 mg/lane protein lysates were separated by using the SDS-PAGE, and the target protein bands were transferred onto the PVDF membranes (Millipore, USA).…”

“…The binding sites of miR-377-3p with hsa_circRNA_103809 and 3' UTR region of GOT1 mRNA were predicted by the online miRDB software (http://mirdb.org/), and validated by using the dual-luciferase reporter gene system, and the detailed experimental procedures had been well documented in the previous literatures [35,36]. Brie y, the targeting sites in hsa_circRNA_103809 and GOT1 were mutated, and named as Mut-circRNA and Mut-GOT1, respectively.…”