MicroRNA-196a-5p is a potential prognostic marker of delayed lymph node metastasis in early-stage tongue squamous cell carcinoma

Abstract

MicroRNAs (miRs) are expected to serve as prognostic tools for cancer. However, many miRs have been reported as prognostic markers of recurrence or metastasis in oral squamous cell carcinoma patients. We aimed to determine the prognostic markers in early-stage tongue squamous cell carcinoma (TSCC). Based on previous studies, we hypothesized that miR-10a, 10b, 196a-5p, 196a-3p, and 196b were prognostic markers and we retrospectively performed miR expression analyses using formalin-fixed paraffin-embedded sections of surgical specimens. Total RNA was isolated from cancer tissues and adjacent normal tissue as control, and samples were collected by laser-capture microdissection. After cDNA synthesis, reverse transcription-quantitative polymerase chain reaction was performed. Statistical analyses for patient clinicopathological characteristics, recurrence/metastasis, and survival rates were performed to discern their relationships with miR expression levels, and the 2−ΔΔCq method was used. miR-196a-5p levels were significantly upregulated in early-stage TSCC, particularly in the lymph node metastasis (LNM) group. The LNM-free survival rate in the low miR-196a-5p ΔΔCq value regulation group was found to be lower than that in the high ΔΔCq value regulation group (P=0.0079). Receiver operating characteristic analysis of ΔΔCq values revealed that miR-196a-5p had a P-value=0.0025, area under the curve=0.740, and a cut-off value=−0.875 for distinguishing LNM. To our knowledge, this is the first study to examine LNM-related miRs in early-stage TSCC as well as miRs and ‘delayed LNM’ in head and neck cancer. miR-196a-5p upregulation may predict delayed LNM. Our data serve as a foundation for future studies to evaluate miR levels and facilitate the prediction of delayed LNM during early-stage TSCC, which prevent metastasis when combined with close follow-up and aggressive adjuvant therapy or elective neck dissection. Moreover, our data will serve as a foundation for future studies to evaluate whether miR-196a-5p can serve as a therapeutic marker for preventing metastasis.

Long non‑coding RNA XIST promotes the proliferation of cardiac fibroblasts and the accumulation of extracellular matrix by sponging microRNA‑155‑5p

“…qPCR amplification parameters were: Annealing at 25˚C for 10 min, extension at 42˚C for 30 min; inactivation at 85˚C for 10 min, after extension of the detection of fluorescence signals, a total of 40 cycles. The 2 -ΔΔCq method ( 16 ) was used to quantify the relative mRNA expression of the target and housekeeping genes. The following primers were used: XIST forward, 5'-ACGCTGCATGTGTCCTTAG-3 and reverse, 5'-GAGCCTCTTATAAGCTGTTTG-3'; miR-155-5p forward, 5'-GAGGGTTAATGCTAATCGTGATAGG-3' and reverse, 5'-GCACAGAATCAACACGACTCACTAT-3'; GAPDH forward, 5'-CTGGGCTACACTGAGCACC-3' and reverse, 5'-AAGTGGTCGTTGAGGGCAATG-3'; and U6 forward, 5'-GAGGGTTAATGCTAATCGTGATAGG-3' and reverse, 5'-GCACAGAATCAACACGACTCACTAT-3'.…”

MicroRNA-532-3p Suppresses Malignant Behaviors of Tongue Squamous Cell Carcinoma via Regulating CCR7

“…MicroRNAs (miRNAs), small non-coding RNAs, play considerable roles in the development of cancer. For instance, miR-196a-5p is a potential prognostic marker of delayed lymph node metastasis in early stage TSCC (Maruyama et al, 2018). It is found that miR-488 is significantly decreased in TSCC tissues and cell lines.…”

miRNA for the assessment of lymph node metastasis in patients with oral squamous cell carcinoma: Systematic review and metanalysis

“…In present review, miRNAs showing statistical significant ( P ≤ .01) correlation with lymph node metastasis includes miRNA 34a, miRNA 196‐5p, miRNA 196‐3p, miRNA 519d, and miRNA 21. All these miRNAs are expressed in tongue squamous cell carcinoma 25‐28 . Also one more common aspect we noticed here is all these miRNAs targets the biomarkers (mRNAs) controlling the epithelial to mesenchymal cell transition (EMT).…”