0001]

The invention relates to prostate cancer including the miRNA expression amount measuring Kit for detection of prostate cancer specific miRNA used to check for prostate cancer incidence in the subjects, and that can be merged into specific nucleic acid that contains the device, and the nucleic acid detection methods.

His innovations.

[0002]

In the organ that produces some of the semen of the male prostate, located in front of the rectum in the bladder. Prostate cancer is a disease this prostate cells repeat the growth disorder. Independent National Cancer Research Center for cancer and, according to the statistics of cancer in Japan to disclose information center 2011 51534 people, the incidence of prostate cancer that affected one a Japanese male 14 people and the incidence in men is fourth cancer site. In addition, deaths from prostate cancer are active 10823, death toll in men of no. 6 cancer site. Also has been in the United States, seven at the rate of one men develop prostate cancer, especially in elderly, at 65 years of age or more in 10 six men will be diagnosed with prostate cancer (non-patent literature 1). In 2014, the United States estimated prostate cancer is up 233000 who is affected by the number and fatality about 29480 people (non-patent literature 1).

[0003]

Prostate cancer progression is provided for non-patent literature 2 spread of the tumor (T1a-c, T2a-c, T3a-b, T4), lymph node metastasis (N0, N1), distant metastases (M0, M1a-c), such as by stage I (T1-T2a/N0/M0), stage II (T2b-c/N0/M0), classified as stage III (T3/N0/M0), stage IV (T4/N0/M0 and N1 and cM1).

[0004]

Many of the prostate cancer progresses relatively slowly in that one of the five-year relative survival rate is almost 100% and the best prognosis cancer (non-patent literature 1). However, during prostate cancer relatively soon and progress and cause a variety of disorders and symptoms, is found in stage 4 metastatic prostate cancer 5-year relative survival rate is 28% and significantly lower (non-patent literature 1).

[0005]

In principle without carry out surgery, radiotherapy, endocrine (hormone therapy), is a special treat prostate cancer treatment, flea treatment, continuing to follow-up and monitor the PSA tumor marker. In particular, in the treatment of early stage prostate cancer and few choices, flea treatment, in addition to external radiotherapy radiation therapy, internal radiation therapy (brachytherapy), radical prostatectomy, cryosurgery, a (non-patent literature 1).

[0006]

PSA test among blood tumor marker in preliminary tests for prostate cancer, as described in the non-patent literature 1, is widely used. Higher PSA readings, rectal diagnosis and percutaneous tissue biopsies were held as a diagnosis when prostate cancer is suspected in addition to transrectal prostate ultrasound examination is carried out. Also, if distant metastasis is suspected, CT, MRI, and bone imaging and scintigraphy examination is done.

[0007]

PSA prostate specific antigen which is produced in the prostate and semen contains the present slightly in the blood. Males usually blood PSA levels have been 4ng/mL less than this threshold has been exceeded, suspected prostate cancer (non-patent literature 1). Serum PSA concentrations is useful from an increase in symptoms of early prostate cancer, that correlate to the progression of the cancer, and widely used. In addition, the United States Cancer Association recommends early detection of prostate cancer, should be admitted PSA test subjects would like screening for prostate cancer and has advised (non-patent literature 1).

[0008]

In the research phase, patent literature 1-is reported to detect prostate cancer by combining the expression level of 3 will be shown, blood and other biological samples in micro RNA (miRNA) or miRNA expression levels and other markers of and.

[0009]

Blood hsa-miR-760, hsa-miR-920, has-miR-887-3p, hsa-miR-486-3p, hsa-miR-663b, hsa-miR-187-5p, hsa-miR-1231, hsa-miR-371a-5p, has-miR-575, hsa-miR-615-5p, hsa-miR-711, hsa-miR-939-5p, hsa-miR-1203, hsa-miR-1225-3p, in the patent literature 1 How to detect a Wilms ' tumor and COPD and other hsa-miR-1225-5p, with hsa-miR-1915-5p, such as prostate cancer and has been shown.

[0010]

Using EpCam vesicles in the blood in the patent literature 2 isolated, have shown how to detect prostate cancer, such as miRNA and hsa-miR-92b-5p contained in the follicles.

[0011]

Is reported to determine prostate cancer by combining the expression of PCA3 gene and expression of miR-141 in the patent literature 3.

Prior art documents

Patent documents

[0012]

Patent literature 1: European patent applications published 2341145 No.
Patent literature 2: international publication No. 2013 / 022995,
Patent literature 3: international publication No. 2010 / 062706,

Non-patent literature

[0013]

Non-patent literature 1: American Cancer Society "Prostate Cancer", in 2013, supervised the p5, 14-26, 32-54, 68-70
Non-patent literature 2: Sobin, L. The TNM Classification of Malignant Tumours, 7th Edition 2010, p. 230-234
Non-patent literature 3: Wolf, AM. From 2010, A Cancer Journal for Clinicians, vol. No. 60 (2), p. 70-98
Non-patent literature 4: Mitchell PS. From 2008, the Proceedings of the National Academy of Sciences of the United Status of America, vol. 105 (30), p. 10513-10518

Summary of the invention

Inventors are trying to solve a problem

[0014]

Challenges of the present invention is a novel prostate cancer to provide how to detect prostate cancer effectively using the heading marker, said marker specifically binding DNA. As a tumor marker test for prostate cancer, the PSA test is widely used. However, PSA test, with 15% 4ng/mL male blood concentrations average biopsy in prostate cancer is known to be judged to be positive, also on the other hand, if men suffering from benign prostate hypertrophy, prostatitis, without cancer, and General man aged in serum PSA concentrations from rising higher false positives also known (non-patent literature 1). Also, even if they accidentally discovered cancer prostate cancer leads to false positives. Lead to overdiagnosis and overtreatment and high false positives in this PSA, prostate cancer without the various side effects of the treatment in recent years held in question (non-patent literature 3). Sensitivity in prostate cancer PSA exact performance depends on the large scale study recruited more than 5,000 participants (non-patent literature 3) and is 20. And only 5% lower, highly malignant prostate cancer also lingering around 51%, meaning as a preoperative tumor marker determination of being.

[0015]

Reports also determine prostate cancer using mRNA is in the research stage blood and other biological samples in micro RNA (miRNA) that have shown below, but none yet practical.

[0016]

Blood hsa-miR-760, hsa-miR-920, has-miR-887-3p, hsa-miR-486-3p, hsa-miR-663b, hsa-miR-187-5p, hsa-miR-1231, hsa-miR-371a-5p, has-miR-575, hsa-miR-615-5p, hsa-miR-711, hsa-miR-939-5p, hsa-miR-1203, hsa-miR-1225-3p, in the patent literature 1 How to detect a Wilms ' tumor and COPD and other hsa-miR-1225-5p, with hsa-miR-1915-5p, such as prostate cancer and has been shown. No direct statements indicated that above miRNA markers are markers of prostate cancer on the other hand, however, that many miRNA described in the patent literature 1, above miRNA marker for prostate cancer is lacking to prove its usefulness as markers.

[0017]

Using EpCam vesicles in the blood in the patent literature 2 isolated, undocumented standards for detecting prostate cancer has been shown to detect prostate cancer miRNA and hsa-miR-92b-5p contained in the follicles, but above miRNA markers in an independent sample group reproduction verification is not too rigid, reliable.

[0018]

Not that easily and accurately determined at single marker is trying to identify prostate cancer in 100% sensitivity and specificity by combining miR-141 and the PCA3 gene specifically in the patent literature 3. Actually, 4 non-patent literature cited in patent literature 3, 4 non-patent literature is prostate cancer miR-141 in serum by discriminant accuracy has been 60% sensitivity and specificity 100%, when the classification has been reported. In addition, urine is a target specimens of the PCA3 test currently used in General, and especially rectal examination has been used after the urine. On the other hand, the miR-141 prostate cancer is determining which samples and blood (serum) as shown above, a combination of two, high sensitivity and specificity of the results should be collected specimens of the two species.

Means for resolving problems

[0019]

To resolve the above issues inventors as a result of the study low reached the finding that can detect prostate cancer significantly by using nucleic acid can be merged specifically to this heading several possible markers for detecting prostate cancer gene from blood samples can be invasive, to perfect the invention.