Methods for diagnosing lung cancer using MicroRNA signatures
The present invention provides novel methods and compositions for the diagnosis and treatment of solid cancers. The invention also provides methods of identifying inhibitors of tumorigenesis.
1. A method of diagnosing whether a subject has, or is at risk for developing, lung cancer, comprising:
receiving a test sample of lung tissue extracted from a subject;
measuring by Northern blot analysis the level of at least miR-205 gene product and at least one additional miR gene product in the test sample, wherein the Northern blot analysis comprises:
extracting nucleic acids from the test sample;
treating the extracted nucleic acids with DNase to remove DNA;
separating RNA molecules in the extracted nucleic acids by gel electrophoresis;
hybridizing the separated RNA molecules to labeled probes; and
detecting and quantifying the hybridization to determine the level of at least miR-205 gene product and at least one additional miR gene product;
comparing the level of at least miR-205 gene product and at least one additional miR gene product in the test sample to a control level of at least one corresponding miR-205 gene product and at least one additional miR gene product in normal lung tissue; and
diagnosing whether the subject has, or is at risk for developing, lung cancer, if the level of the at least miR-205 gene product and at least one additional miR gene product in the test sample from the subject is greater than the control level of at least one miR-205 gene product and at least one corresponding additional miR gene product.
2. A method of claim 1 , wherein the at least one additional miR gene product is miR-21.
3. A method of claim 1 , wherein the at least one additional miR gene product is miR-200b.
4. A method of claim 1 , wherein the at least one additional miR gene product is miR-9-1.
5. A method of claim 1 , wherein the at least one additional miR gene product is miR-210.
6. A method of claim 1 , wherein the at least one additional miR gene product is miR-148.
7. A method of claim 1 , which further comprises: comparing the level of at least one additional miR gene product in a test sample from the subject to a control level of the miR gene product, wherein the at least one additional miR gene product is selected from the group consisting of: miR-21; miR-200b; miR-9-1; miR-210; miR-148; miR-141; miR-132; miR-215; miR-128b; let-7g; miR-16-2; miR-129-1/2 prec; miR-126; miR-142-as; miR-30d; miR-30a-5p; miR-7-2; miR-199a-1; miR-127; miR-34a prec; miR-34a; miR-136; miR-202; miR-196-2; miR-199a-2; let-7a-2; miR-124a-1; miR-149; miR-17-5p; miR-196-1 prec; miR-10a; miR-99b prec; miR-196-1; miR-199b; miR-195; and miR-155.
8. A method of claim 1 , which further comprises: comparing the level of at least one additional miR gene product in a test sample from the subject to a control level of the miR gene product, wherein the at least one additional miR gene product is selected from the group consisting of: miR-21; miR-17-5p; miR-29b-2; miR-223; miR-128b; miR-199a-1; miR-24-1; miR-24-2; miR-146; miR-155; miR-181b-1; miR-20a; miR-107; miR-32; miR-92-2; miR-214; miR-30c; miR-25; miR-221; and miR-106a.