CN102409091A - Method for using prostate secretory protein 94/57 as biomarker for early clinical diagnosis of ovarian cancer and breast cancer - Google Patents
Method for using prostate secretory protein 94/57 as biomarker for early clinical diagnosis of ovarian cancer and breast cancer Download PDFInfo
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- CN102409091A CN102409091A CN2011103370064A CN201110337006A CN102409091A CN 102409091 A CN102409091 A CN 102409091A CN 2011103370064 A CN2011103370064 A CN 2011103370064A CN 201110337006 A CN201110337006 A CN 201110337006A CN 102409091 A CN102409091 A CN 102409091A
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- 108091058545 Secretory proteins Proteins 0.000 title claims abstract description 46
- 102000040739 Secretory proteins Human genes 0.000 title claims abstract description 46
- 206010006187 Breast cancer Diseases 0.000 title claims abstract description 25
- 208000026310 Breast neoplasm Diseases 0.000 title claims abstract description 25
- 206010033128 Ovarian cancer Diseases 0.000 title claims abstract description 24
- 206010061535 Ovarian neoplasm Diseases 0.000 title claims abstract description 24
- 238000000034 method Methods 0.000 title claims abstract description 13
- 210000002307 prostate Anatomy 0.000 title claims description 47
- 239000000090 biomarker Substances 0.000 title abstract description 12
- 238000003759 clinical diagnosis Methods 0.000 title description 4
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 40
- 230000014509 gene expression Effects 0.000 claims abstract description 17
- 101150003382 57 gene Proteins 0.000 claims description 28
- 102000004169 proteins and genes Human genes 0.000 claims description 28
- 108020004999 messenger RNA Proteins 0.000 claims description 20
- 210000001519 tissue Anatomy 0.000 claims description 20
- 210000004369 blood Anatomy 0.000 claims description 12
- 239000008280 blood Substances 0.000 claims description 12
- 238000012360 testing method Methods 0.000 claims description 7
- 238000001514 detection method Methods 0.000 claims description 6
- 238000003745 diagnosis Methods 0.000 claims description 5
- 239000000523 sample Substances 0.000 claims 6
- 210000001124 body fluid Anatomy 0.000 claims 4
- 239000010839 body fluid Substances 0.000 claims 4
- 239000003153 chemical reaction reagent Substances 0.000 claims 4
- 210000002700 urine Anatomy 0.000 claims 4
- -1 urine Substances 0.000 claims 4
- 230000002068 genetic effect Effects 0.000 claims 3
- 238000002965 ELISA Methods 0.000 claims 2
- 238000004458 analytical method Methods 0.000 claims 2
- 238000003364 immunohistochemistry Methods 0.000 claims 2
- 239000000126 substance Substances 0.000 claims 2
- 201000011510 cancer Diseases 0.000 abstract description 14
- 238000013399 early diagnosis Methods 0.000 abstract description 5
- 239000013610 patient sample Substances 0.000 abstract 1
- 206010028980 Neoplasm Diseases 0.000 description 17
- 201000001514 prostate carcinoma Diseases 0.000 description 6
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 238000005336 cracking Methods 0.000 description 4
- 210000001672 ovary Anatomy 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 208000024891 symptom Diseases 0.000 description 4
- 238000001262 western blot Methods 0.000 description 4
- 230000002547 anomalous effect Effects 0.000 description 3
- 238000011529 RT qPCR Methods 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 238000010298 pulverizing process Methods 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 206010027336 Menstruation delayed Diseases 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 201000008275 breast carcinoma Diseases 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 208000023958 prostate neoplasm Diseases 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
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Abstract
The invention provides a method for clinical early diagnosis of ovarian cancer and breast cancer. The method uses a human self gene-prostatic secretory protein 94/57 as a biomarker, and can preliminarily determine whether a patient has malignant tumor by detecting the expression quantity, existence or nonexistence of the gene in a patient sample.
Description
[invention field]
Biotechnology and medical field are specially the early stage clinical detection method that relates to a kind of indication, diagnosis of ovarian cancer, mammary cancer under the present invention.
[technical background]
Cancer is to threaten one of the most serious disease of human health.Although obtained a series of progress at its treatment, diagnostic field in recent years, however the cancer high a kind of disease that still is mortality ratio.Early diagnostic rate is low, and the forfeiture best occasion for the treatment is to cause cancer weak curative effect and the high major cause of dead rate.Therefore improve the early diagnostic rate of cancer, and then and early treatment be one of effective means of defeating this disease.Most of cancer patients does not have tangible clinical symptom in early days in disease, in case manifest symptom appears then in having got into, late period.Therefore the early diagnosis of cancer, the especially diagnosis before manifest symptom occurring have very important significance.
Research shows most of cancer is because the unconventionality expression of human body specific gene or close unusually causes, and wherein the ANOMALOUS VARIATIONS of portion gene is the very early time incident that cancer takes place, and it can produce symptom patient and take place before.Therefore the ANOMALOUS VARIATIONS of these specific genes can be used as the biomarker of early diagnosis of cancer.Use these affinity tags, combine conventional methods for clinical diagnosis diagnosed disease fast and effectively again, thus for the cancer patients's and early treatment win the best moment.
Prostate gland secretory protein 94 is one and contains 94 amino acid whose small proteins (it also has a homogenic isomer, contains 57 amino acid), mainly by the prostate gland secretion, is one of three the abundantest large proteins of prostate gland secretion.Some researchs show that this proteic expression amount significantly reduces in the carcinoma of prostate patient; From early stage patient to, end-stage patients; The trend that the patient tapers off; To such an extent as among the patient, this proteic expression amount completely dissolve is so this albumen is a potential biomarker of carcinoma of prostate patient clinical diagnosis late.Simultaneously, some researchs show that also this albumen has the function that suppresses prostate tumor cells growth, kill cancer cell.In addition; Show according to up-to-date result of study; This proteic gene of encoding is the sensitive gene of a carcinoma of prostate, has a single base mutation at the control region of this gene promoter; This sudden change can cause its expressing quantity to descend, thereby makes the individuality that carries sudden change become the susceptible patient of prostate cancer card.And correlative study shows that many carcinoma of prostate patients carry this single base mutation.Therefore this gene has important latent effect in generation, development and the therapeutic process of carcinoma of prostate, and is the biomarker that significant a kind of potential detects carcinoma of prostate.Because the ANOMALOUS VARIATIONS of most gene has the cancer types specificity, so whether this gene have biological function in the generating process of other types cancer, also do not carry out relevant research.
[summary of the invention]
The present invention shows through experiment: the expression amount of prostate gland secretory protein 94/57 significantly raises in the sample of ovarian cancer patients, and its expression amount (comprising mRNAs and albumen) will exceed 2 to 5 times than normal people expression amount.Especially in early days in patient's sample.Infer that thus prostate gland secretory protein 94/57 significantly raises in the ovarian cancer early stage patient, it can be used as a kind of indication, diagnosis of ovarian cancer early detection potential biomarker.
The present invention shows through test: the expression amount of prostate gland secretory protein 94/57 (comprising mRNAs and albumen) significantly reduces in patient with breast cancer's sample, to such an extent as to even completely dissolve detect less than.Especially in early days in patient's sample.Infer that in view of the above prostate gland secretory protein 94/57 reduces in the mammary cancer early stage patient unusually, it can be used as a kind of indication, diagnosing mammary cancer early detection potential biomarker.
[concrete real-time mode]
Through following embodiment the present invention is described further.
One: the mRNAs of prostate gland secretory protein 94/57 gene and protein level significantly raise in the ovarian cancer early stage patient.
1) the mRNAs level of prostate gland secretory protein 94/57 gene is suffered from significantly rising among the early stage person in ovarian cancer.Get the tumor tissues appearance of ovarian cancer patients and normal people's ovary tissue appearance and handle (extracting mRNAs, reverse transcription), detect the mRNAs level of prostate gland secretory protein 94/57 gene then through real-time quantitative-PCR.The result shows: in the ovary tissue sample, the prostate gland secretory protein 94/57mRNAs level of ovarian cancer early stage patient is normal people about 6 times (seeing table 1).
2) protein level of prostate gland secretory protein 94/57 gene significantly raises in the ovarian cancer early stage patient.
A) get the tumor tissues appearance of ovarian cancer patients and normal people's ovary tissue appearance and handle (pulverizing, cracking whole protein), detect the protein level of prostate gland secretory protein 94/57 gene then through Western Blot.The result shows: in the ovary tissue sample, the protein level of the prostate gland secretory protein 94/57 of ovarian cancer early stage patient is normal people's about 3.2 times (seeing table 2).
B) get the blood sample of ovarian cancer patients and normal people's blood sample and handle (separation of serum, cracking whole protein), detect the protein level of prostate gland secretory protein 94/57 gene then through Western Blot.The result shows: in blood sample, the protein level of the prostate gland secretory protein 94/57 of ovarian cancer early stage patient is normal people's about 2 times (seeing table 3).
Table 1: the comparison of the mRNAs level of prostate gland secretory protein 94/57 gene in ovarian cancer patient and health adult tissue's appearance
Table 2: the comparison of the protein level of prostate gland secretory protein 94/57 gene in ovarian cancer patient and health adult tissue's appearance
Table 3: the comparison of the protein level of prostate gland secretory protein 94/57 gene in ovarian cancer patient and normal people's blood sample
Two: the mRNAs of prostate gland secretory protein 94/57 gene and protein level significantly reduce in the mammary cancer early stage patient.
1) the mRNAs level of prostate gland secretory protein 94/57 gene significantly reduces in the patient with breast cancer.Get patient with breast cancer's tumor tissues appearance and normal people's mammary tissue appearance and handle (extracting mRNAs, converse transcribing), detect the mRNAs level of prostate gland secretory protein 94/57 gene then through real-time quantitative-PCR.The result shows: in the mammary tissue sample, patient with breast cancer's prostate gland secretory protein 94/57mRNAs level is about 1/5th (seeing table 4) of normal people.
2) protein level of prostate gland secretory protein 94/57 gene significantly reduces in the mammary cancer early stage patient.
A) get patient with breast cancer's tumor tissues appearance and normal people's mammary tissue appearance and handle (pulverizing, cracking whole protein), detect the protein level of prostate gland secretory protein 94/57 gene then through Western Blot.The result shows: in the mammary tissue sample, the protein level of the prostate gland secretory protein 94/57 of mammary cancer early stage patient is normal people's 1/3rd (seeing table 5).
B) get patient with breast cancer's blood sample and normal people's blood sample and handle (separation of serum, cracking whole protein), detect the protein level of prostate gland secretory protein 94/57 gene then through Western Blot.The result shows: in blood sample, the protein level of the prostate gland secretory protein 94/57 of mammary cancer early stage patient is extremely low, to such an extent as to detect less than (seeing table 6).
Table 4: the comparison of the mRNAs level of prostate gland secretory protein 94/57 gene in mammary cancer patient and health adult tissue's appearance
Table 5: the comparison of the protein level of prostate gland secretory protein 94/57 gene in mammary cancer patient and health adult tissue's appearance
Table 6: the comparison of the protein level of prostate gland secretory protein 94/57 gene in mammary cancer patient and normal people's blood sample
Above-mentioned test-results of the present invention shows:
1, prostate gland secretory protein 94/57 significantly raises in HOC's early stage patient, and it can be used as one of biomarker or biomarker of the clinical early diagnosis of ovarian cancer.
2, prostate gland secretory protein 94/57 significantly reduces in the human breast carcinoma early stage patient, and it can be used as one of biomarker or biomarker of the clinical early diagnosis of mammary cancer.
Reference
1.Nossov，V.，et?al.，The?early?detection?of?ovarian?cancer：from?traditional?methods?to?proteomics.Can?we?really?do?better?than?serum?CA-125?Am?J?Obstet?Gynecol，2008.199(3)：p.215-23.
2.Bast，R.C.，Jr.，et?al.，Prevention?and?early?detection?of?ovarian?cancer：mission?impossible?Recent?Results?Cancer?Res，2007.174：p.91-100.
3.Badgwell，D.and?R.C.Bast，Jr.，Early?detection?of?ovarian?cancer.Dis?Markers，2007.23(5-6)：p.397-410.
4.Lilja，H.and?P.A.Abrahamsson，Three?predominant?proteins?secreted?by?the?human?prostate?gland.Prostate，1988.12(1)：p.29-38.
5.Weiber，H.，et?al.，Beta?microseminoprotein?is?not?a?prostate-specific?protein.Its?identification?in?mucous?glands?and?secretions.Am?J?Pathol，1990.137(3)：p.593-603.
6.Baijal-Gupta，M.，et?al.，Prostatic?secretory?protein(PSP94)expression?in?human?female?reproductive?tissues，breast?and?in?endometrial?cancer?cell?lines.J?Endocrinol，2000.165(2)：p.425-33.
7.Whitaker，H.C.，et?al.，The?potential?value?of?microseminoprotein-beta?as?a?prostate?cancer?biomarker?and?therapeutic?target.Prostate.70(3)：p.333-40.
8.Thomas，G.，et?al.，Multiple?loci?identified?in?a?genome-wide?association?study?of?prostate?cancer.Nat?Genet，2008.40(3)：p.310-5.
9.Eeles，R.A.，et?al.，Multiple?newly?identified?loci?associated?with?prostate?cancer?susceptibility.Nat?Genet，2008.40(3)：p.316-21.
10.Lou，H.，et?al.，Fine?mapping?and?functional?analysis?of?a?common?variant?in?MSMB?on?chromosome?10q11.2?associated?with?prostate?cancer?susceptibility.Proc?Natl?Acad?Sci?U?S?A，2009.106(19)：p.7933-8.
11.Xu，B.，et?al.，A?functional?polymorphism?in?MSMB?gene?promoter?is?associated?with?prostate?cancer?risk?and?serum?MSMB?expression.Prostate.70(10)：p.1146-52.
12.Shukeir，N.，et?al.，A?synthetic?15-mer?peptide(PCK3145)derived?from?prostate?secretory?protein?can?reduce?tumor?growth，experimental?skeletal?metastases，and?malignancy-associated?hypercalcemia.Cancer?Res，2004.64(15)：p.5370-7.
13.Hawkins，R.E.，et?al.，Safety?and?tolerability?of?PCK3145，a?synthetic?peptide?derived?from?prostate?secretory?protein?94(PSP94)in?metastatic?hormone-refractory?prostate?cancer.Clin?Prostate?Cancer，2005.4(2)：p.91-9.
14.Shukeir，N.，et?al.，Prostate?secretory?protein?PSP-94?decreases?tumor?growth?and?hypercalcemia?of?malignancy?in?a?syngenic?in?vivo?model?of?prostate?cancer.Cancer?Res，2003.63(9)：p.2072-8.
Claims (6)
1. the detection method of an indication, diagnosis of ovarian cancer, this method are used or are comprised use:
A) detecting prostate gland secretory protein 94/57 gene has, does not have expressions in testing sample (blood, urine, body fluid, tissue appearance etc.), comprising the mRNA of this gene of detection perhaps proteic having or not;
B) detect the expression amount of prostate gland secretory protein 94/57 gene in testing sample (blood, urine, body fluid, tissue appearance etc.), comprising mRNAs that detects this gene or proteic expression amount;
, genetic expression indicates that then ovarian cancer produces when surpassing setting threshold.
2. according to the said method of claim 1, it comprises use:
A) RT-PCR (converse transcribing-polymerase chain amplified reaction), quantitatively-PCR, methods such as gene chip detect having or not or what of prostate gland secretory protein 94/57 gene mRNA s;
B) methods such as Western, ELISA, immunohistochemistry (immunohistichemistry), chromatogram analysis of protein detect having or not or what of prostate gland secretory protein 94/57 gene protein.
3. the test kit of an indication, diagnosis of ovarian cancer, its characteristic comprises use:
A) detect having or not or what reagent such as primer or probe of prostate gland secretory protein 94/57 gene mRNA s;
B) detect having or not or what reagent such as antibody of prostate gland secretory protein 94/57 gene protein;
C) comprise that ovarian cancer is positive in article, negative standard substance with reference to article.
4. the detection method of an indication, diagnosing mammary cancer, this method are used or are comprised use:
A) detect prostate gland secretory protein 94/57 genetic expression having or not in testing sample (blood, urine, body fluid, tissue appearance etc.), comprising mRNA that detects this gene or proteic having or not;
B) detect the expression amount of prostate gland secretory protein 94/57 genetic expression in testing sample (blood, urine, body fluid, tissue appearance etc.), comprising mRNAs that detects this gene or proteic expression amount;
, then indicates gene expression amount that mammary cancer produces when being lower than setting threshold.
5. according to the said method of claim 4, its use comprises:
A) RT-PCR (converse transcribing-polymerase chain amplified reaction), quantitatively-PCR, methods such as gene chip detect having or not or what of prostate gland secretory protein 94/57 gene mRNA s;
B) methods such as Western, ELISA, immunohistochemistry (immunohistichemistry), chromatogram analysis of protein detect having or not or what of prostate gland secretory protein 94/57 gene protein.
6. the test kit of an indication, diagnosing mammary cancer, its characteristic comprises use:
A) detect having or not or what reagent such as primer or probe of prostate gland secretory protein 94/57 gene mRNA s;
B) detect having or not or what reagent such as antibody of prostate gland secretory protein 94/57 gene protein;
C) comprise that mammary cancer is positive in article, negative standard substance with reference to article.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1537170A (en) * | 2000-11-20 | 2004-10-13 | ����ҽѧԺ | Method and apparatus for quantitative detection of prostate specific membrane antigens and other prostate markers |
| WO2006133560A1 (en) * | 2005-06-17 | 2006-12-21 | Ambrilia Biopharma Inc. | Psp94 diagnostic reagents and assays |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1537170A (en) * | 2000-11-20 | 2004-10-13 | ����ҽѧԺ | Method and apparatus for quantitative detection of prostate specific membrane antigens and other prostate markers |
| WO2006133560A1 (en) * | 2005-06-17 | 2006-12-21 | Ambrilia Biopharma Inc. | Psp94 diagnostic reagents and assays |
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Application publication date: 20120411 |