miRNA fingerprint in the diagnosis of lung cancer
The present invention provides novel methods for diagnosing diseases based on the determination of specific miRNAs that have altered expression levels in disease states compared to healthy controls.
1. A method of detecting miRNA, comprising
(a) extracting total intracellular RNA from all blood cells obtained from a whole blood sample taken from a patient, and
(b) determining in said total intracellular RNA an expression profile of an miRNA set consisting of:
(i) hsa-miR-423-5p, hsa-miR-126, and hsa-let-7i;
(ii) hsa-miR-423-5p, hsa-miR-126, hsa-let-7i, and hsa-let-7d;
(iii) hsa-miR-423-5p, hsa-miR-126, hsa-let-7i, hsa-let-7d, and hsa-miR-22; or
(iv) hsa-miR-423-5p, hsa-miR-126, hsa-let-7i, hsa-let-7d, hsa-miR-22, hsa-miR-15a, hsa-miR-98, hsa-miR-19a, hsa-miR-574-5p, and hsa-miR-324-3p.
2. The method of claim 1 , wherein the patient is suspected of suffering from lung cancer.
3. The method of claim 1 , wherein the lung cancer is selected from the group consisting of lung carcinoid, lung pleural mesothelioma, non-small cell lung carcinoma and lung squamous cell carcinoma.
4. The method of claim 3 , wherein the lung cancer is non-small cell lung carcinoma.
5. The method of claim 1 , wherein step (b) comprises quantitative or semiquantitative detection of said miRNAs.
6. The method of claim 1 , wherein determining said expression profile comprises nucleic acid amplification performed using real-time polymerase chain reaction (PCR).
7. The method of claim 6 , wherein determining the expression profile using real-time PCR comprises:
(1) reverse transcribing the total intracellular RNA extracted from said blood cells into cDNA, and
(2) quantifying the cDNA, thereby determining the expression profile of said miRNAs.
8. The method of claim 1 , wherein said blood cells comprise a mixture of erythrocytes, leukocytes, and thrombocytes.
9. The method of claim 1 , wherein the expression profile of hsa-miR-423-5p, hsa-miR-126, hsa-let-7i, and hsa-let-7d is determined.