Herein, we develop an isothermal miRNA detection platform based on the highly efficient, multiple primer-mediated rolling circle amplification method coupled with a graphene oxide-based fluorescence (MPRCA-GO) assay, using lung cancer-associated miRNAs (miR-21 and miR-210) and a reference miRNA (miR-16) as model targets.
Next, transfection of miR-210 mimics in lung cancer cells induced significant increase in cell proliferation, and transfection of miR-210 inhibitors lead to inhibited cell proliferation.
MiR-210 expression level in the diagnosis of COPD complicating lung cancer had the sensitivity of 76.8%, specificity of 72.3%, accuracy of 0.73, 95% CI of 0.63-0.85 and critical value of 0.1825.
Detailed analysis revealed significant change in expression of miR-15a/16, miR-34a, miR-126, and miR-210 in NSCLC tumor samples indicating involvement of these miRNAs in lung cancer pathogenesis. miR-210 demonstrated the most consistent increase in tumor tissues between different patients, suggesting its potential significance for NSCLC.
Furthermore, combined quantification of miR-31 and miR-210 copy number by using digital PCR in sputum of the cases and controls provided 65.71 % sensitivity and 85.00 % specificity for lung cancer diagnosis.
From the miRNAs, a logistic regression model was built on the basis of miR-31 and miR-210, both of which had the best prediction for lung cancer, producing an area under receiver operating characteristic curve of 0.83.
These include miR-301, miR-183/96/182, miR-126, and miR-223, which are microRNAs deregulated in other tumor types as well; and other miRNAs, such as miR-374 and miR-210, not previously reported in association with lung cancer.