Methods for diagnosing breast, colon, lung, pancreatic and prostate cancer using miR-21 and miR-17-5p

1. A method of diagnosing whether a subject has, or is at risk for developing, a solid cancer, comprising:

receiving a test sample of breast, colon, lung, pancreatic, or prostate tissue extracted from a subject;

measuring by Northern blot analysis the level of at least miR-21 and miR-17-5p gene products 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-21 and miR-17-5p gene products;

comparing the level of at least miR-21 and miR-17-5p gene products in the test sample to a control level of at least miR-21 and miR-17-5p gene products in a control sample of normal corresponding tissue; and

diagnosing whether the subject has, or is at risk for developing, a solid cancer other than stomach cancer if the level of at least miR-21 and miR-17-5p gene products in the test sample from the subject is greater than the level of miR-21 and miR-17-5p gene products of the control;

wherein:

elevated levels of miR-21 and miR-17-5p gene products in breast tissue from the subject as compared to the control indicate the subject has breast cancer;

elevated levels of miR-21 and miR-17-5p gene products in colon tissue from the subject as compared to the control indicate the subject has colon cancer;

elevated levels of miR-21 and miR-17-5p gene products in lung tissue from the subject as compared to the control indicate the subject has lung cancer;

elevated levels of miR-21 and miR-17-5p gene products in pancreatic tissue from the subject as compared to the control indicate the subject has pancreatic cancer; and

elevated levels of miR-21 and miR-17-5p gene products in prostate tissue from the subject as compared to the control indicate the subject has prostate cancer.

2. A method of diagnosing whether a subject has, or is at risk for developing, a solid cancer, comprising:

receiving a test sample of colon or pancreatic tissue extracted from a subject;

reverse transcribing at least miR-21 and miR-17-5p from the test sample to provide at least miR-21 and miR-17-5p target oligonucleotides;

hybridizing at least the miR-21 and miR-17-5p target oligodeoxynucleotides to a microarray comprising miRNA-specific probe oligonucleotides covalently attached to a polymeric matrix, wherein the probe oligonucleotides include at least miR-21 and miR-17-5p RNA specific probe oligonucleotides, to provide a hybridization profile for the test sample;

comparing the test sample hybridization profile to at least one control hybridization profile from normal corresponding tissue; and

diagnosing whether the subject has, or is at risk for developing, a solid cancer selected from the group consisting of: colon cancer and pancreatic cancer, if the level of at least miR-21 and miR-17-5p gene products in the test sample from the subject is greater than the level of miR-21 and miR-17-5p gene products of the control;

wherein:

elevated levels of miR-21 and miR-17-5p gene products in colon tissue from the subject as compared to the control indicate the subject has colon cancer; and

elevated levels of miR-21 and miR-17-5p gene products in pancreatic tissue from the subject as compared to the control indicate the subject has pancreatic cancer.

3. The 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-191; 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.

4. The method of claim 1 , which further comprises comparing the level of at least two additional miR gene products in a test sample from the subject to a control level of the corresponding miR gene products, wherein the at least two additional miR gene products are selected from the group consisting of: miR-191; 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.

5. The method of claim 1 , which further comprises comparing the level of at least three additional miR gene products in a test sample from the subject to a control level of the corresponding miR gene products, wherein the at least three additional miR gene products are selected from the group consisting of: miR-191; 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.

6. The method of claim 1 , wherein the solid cancer is breast cancer and 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-29b-2; miR-146; miR-125b-2; miR-125b-1; miR-10b; miR-145; miR-181a; miR-140; miR-213; miR-29a prec; miR-181b-1; miR-199b; miR-29b-1; miR-130a; miR-155; let-7a-2; miR-205; miR-29c; miR-224; miR-100; miR-31; miR-30c; miR-210; miR-122a; and miR-16-2.

7. The method of claim 1 , wherein the solid cancer is colon cancer and 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-24-1; miR-29b-2; miR-20a; miR-10a; miR-32; miR-203; miR-106a; miR-30c; miR-223; miR-126; miR-128b; miR-24-2; miR-99b prec; miR-155; miR-213; miR-150; miR-107; miR-191; miR-221; and miR-9-3.

8. The method of claim 1 , wherein the solid cancer is lung cancer and 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-205; 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-196-1 prec; miR-10a; miR-99b prec; miR-196-1; miR-199b; miR-191; miR-195; and miR-155.

9. The method of claim 1 , wherein the solid cancer is pancreas cancer and 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-103-2; miR-103-1; miR-24-2; miR-107; miR-100; miR-125b-2; miR-125b-1; miR-24-1; miR-191; miR-23a; miR-26a-1; miR-125a; miR-130a; miR-26b; miR-145; miR-221; miR-126; miR-16-2; miR-146; miR-214; miR-99b; miR-128b; miR-155; miR-29b-2; miR-29a; miR-25; miR-16-1; miR-99a; miR-224; miR-30d; miR-92-2; miR-199a-1; miR-223; miR-29c; miR-30b; miR-129-1/2; miR-197; miR-30c; miR-7-1; miR-93-1; miR-140; miR-30a-5p; miR-132; miR-181b-1; miR-152 prec; miR-23b; miR-20a; miR-222; miR-27a; miR-92-1; miR-129-1/2 prec; miR-150; miR-32; miR-106a; and miR-29b-1.

10. The method of claim 1 , wherein the solid cancer is prostate cancer and 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: let-7d; miR-128a prec; miR-195; miR-203; let-7a-2 prec; miR-34a; miR-20a; miR-218-2; miR-29a; miR-25; miR-95; miR-197; miR-135-2; miR-187; miR-196-1; miR-148; miR-191; let-7i; miR-198; miR-199a-2; miR-30c; miR-92-2; miR-146; miR-181b-1 prec; miR-32; miR-206; miR-184 prec; miR-29a prec; miR-29b-2; miR-149; miR-181b-1; miR-196-1; miR-93-1; miR-223; miR-16-1; miR-101-1 prec; miR-124a-1; miR-26a-1; miR-214; miR-27a; miR-24-1; miR-106a; and miR-199a-1.