JP2002538433A - Staging of prostate cancer - Google Patents

Abstract

  (57) [Summary] The present invention relates to a method for staging prostate cancer, i.e., discriminating between organ-localized prostate cancer (PCa) and non-organ-localized PCa in a patient, wherein human glandular kallikrein 2 (hK2) ) And optionally prostate specific antigen (PSA). In this method, hK2 is used as a marker to classify patients with organ-localized PCa from patients with organ-non-localized PCa. In addition, the present invention provides a method for grading prostate cancer, i.e., for distinguishing patients with aggressively progressing prostate cancer (PCa) from patients with less aggressively progressing PCa. For the method, human glandular kallikrein 2 (hK2) in a patient's body fluid is now determined. In this method, hK2 is used alone as the marker.

Description

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to a method for differentiating organ-localized prostate cancer (PCa) in a patient from non-organ-localized PCa, ie, for staging a patient's PCa. Furthermore, the invention relates to a method for distinguishing patients with aggressively progressing PCa from patients with less aggressively progressing PCa, i.e. for grading the patient's PCa prior to surgery.

INTRODUCTION AND BACKGROUND Publications and other materials used to elucidate the background of the invention and, in particular, to provide additional details regarding the practice of the invention, are incorporated by reference.

[0003] Prostate cancer (PCa) is the most commonly diagnosed cancer in men, and mortality from PCa is second only to lung tumors ¹ . With the advent of the prostate-specific antigen (PSA) ^{2 in} 1971 and its introduction into clinical use in 1979, prostate-specific antigen has emerged as the most important tumor marker in urological professionals ^4,5 . Areas of much value for the clinical application is the follow-up after surgery after radical prostate removal, where, with sufficient organ specificity for all implementation purposes ^6, recurrent disease Evidence may be based solely on the re-emergence of PSA in serum ^6-11 .

[0004] Prostate cancer susceptibility and despite its localized due to the lack of specificity ^{5,9,12,1 3,} its use has also been established in the diagnosis of prostate cancer. However, the lack of sensitivity and specificity suggests that PSA could be used to enhance the clinical utility of this tumor marker.
Parameters, especially PSA rate ¹⁴ , PSA density ¹⁵ , metastatic PSA density ¹⁶ , age-specific PSA range ^17, and free PSA for total PSA
Resulting in the production ratio (% fPSA) ^{18, 19, 20} of.

[0005] The application of PSA in preoperative staging of prostate cancer has shown that serum PSA levels correlate with tumor volume, clinical and pathological stage progression ^7,9,13 . On the other hand, individual criteria have shown that PSA levels alone are not specific enough to allow accurate prediction of the final pathological stage ^11,12 . The most effective approach in the treatment of prostate cancer can be performed when the tumor is further localized. Information can be obtained from the results of a regular six-part biopsy. However, serum markers for more accurate staging of the prostate will provide important and more readily available new information.

At present, closely related to the same family of enzymes, the so-called serine proteases
Protein, a novel serum marker of prostate origin, human glandular kallikrein
2 (hK2) is a potential marker for benign and malignant differentiation for prostate tissue
Has become apparent^21,22,23. hK2 gene is 80% of PSA gene
There is homology. Recent studies have shown that both hK2 and PSA mRNA
Suggested to be found exclusively in^24,25,26. They are also Andro
Shares the features of gene-regulated expression^25,27 . Finally, hK2 opens pro-PSA
Is an enzyme that cleaves and thus activates it into its enzymatically active form ²⁸ . Less common use for human glandular kallikrein 3 for prostate specific antigen
The term foretells a link between the two enzymes.

[0007] From a biochemical point of view, the close homology of both proteins required the design of specific monoclonal antibodies that did not cross-react with other kallikreins as much as possible. hK
Recent epitope mapping studies of ^PSA2 and PSA have ^29,30 , varying degrees of anti-PS
³¹ showing the cross-reactivity with A antibodies and hK2. Based on this information, different designs of immunoassays were generated for specific measurement of hK2 ²⁹ .

[0008] Clinically, serum concentrations of hK2 are 4-10 ng / ml (diagnostic gray zone) in the detection of prostate cancer in patients with a total PSA of ³² , and hK2 (and PSA) in radical prostatectomy specimens. ) Has been used in an attempt to improve the measurement of cytoplasmic expression ²² ).

[0009] Clinically, inadequate staging of prostate tumors selects treatment with therapeutic intent
The main problem is when. Undergoing radical prostatectomy, clinically limited organs
Of localized prostate tumors, 26-43% exhibit extra-articular disease^{35,3 6} . 30-80% of these will progress to progressive disease within 10 years^{37 , 38} . Therefore, new diagnostic tools to reduce this inadequate staging are needed.
There is a clear need. Prostate in addition to unsatisfactory clinical T stage
The prognosis of adenocarcinoma also depends on the histologic grade of the tumor^38,39 And
This is done before surgery with a sextant biopsy
And often unreliable. In addition, increased serum PSA levels are associated with more advanced pathology.
Does not properly reflect the radiological grade, especially in the middle area of PSA ^12,40,41 .

Objects and Summary of the Invention It is an object of the invention to provide a method for staging a patient's PCa.

In an initial study, we determined whether hK2 levels differed in various pathological stagings, and furthermore whether hK2 levels differed in patients with organ-localized and non-organ-localized prostate cancer. To evaluate, we focused our attention on the concentration of hK2 in 68 radical prostatectomy patients. PSA is
Since it is impossible to certainly predict gun localized organ ^{11, 12} for the individual patient, this ability in biochemical staging of preoperative adenocarcinoma of the prostate, the new serum markers Could be an important feature.

Another object of the present invention is to provide a method for assessing the grade of PCa of a patient.

[0013] Human glandular kallikrein 2 (hK2) binds to prostate specific antigen (PSA)
It has 0% structural identity and is secreted by the same prostate epithelial cells. Although increasing with pathological staging, serum PSA is not clinically useful in assessing aggressiveness of prostate cancer in individuals. hK2 itself has a different PS
A study was undertaken to evaluate whether to allow differentiation of well-, moderately- and poorly differentiated prostate cancer compared to type A.

Thus, in accordance with one aspect, the present invention relates to a method for staging prostate cancer, ie, for distinguishing a patient's organ-localized PCa from an organ-non-localized PCa, wherein the patient's human Fluid concentrations of glandular kallikrein 2 (hK2) and optionally also prostate specific antigen (PSA) are determined. According to the present invention, hK2 is used as a marker to distinguish patients with organ-localized PCa from patients with organ-non-localized PCa.

In accordance with another aspect, the present invention is directed to a method for grading prostate cancer, ie, reducing a patient with aggressively advancing PCa to a less aggressive PCa.
Wherein the body fluid concentration of human glandular kallikrein 2 (hK2) in the patient is determined. According to the invention, hK2 is used alone as said marker.

DETAILED DESCRIPTION OF THE INVENTION According to a preferred embodiment, hK2 is used alone as a staging marker. hK2 and PSA (where PSA is free PSA, complex PSA or total PSA
A) can alternatively be used as said marker. In the latter case, the marker is preferably the hK2 × total PSA / free PSA algorithm.

The grading method preferably comprises: i) distinguishing between patients with well-differentiated and moderately differentiated PCa on the one hand and patients with poorly differentiated PCa on the other hand, or ii) patients with moderately differentiated PCa on the other hand; On the other hand, either discrimination from patients with poorly differentiated PCa.

[0018] The malignancy classification method has a total P in the range of 1 to 20 ng / ml, particularly in the range of 3 to 15 ng / ml.
It is particularly useful in patients with SA.

The detection of PSA in serum, both conjugated and unconjugated, has begun as an established part of the pre- and post-treatment evaluation of prostate cancer. Once a histological diagnosis of prostate cancer has been made and the patient's assessment suggests clinically localized prostate cancer, radical prostatectomy may be applied to a therapeutic attempt for the patient It will be the standard for treatment.
However, studies could show that 40-60% of patients had capsule invasion of the cancer. Prognosis is closely related to pathological staging, especially in terms of survival unrelated to PSA.

Patients with cancers that have pathologically unrestricted organs may have 90
% Of PSA-independent survival and consequently possible cure rates are achieved, while capsule invasion (T3a or higher pathological staging) is associated with PSA-independent survival And surprisingly reduce the healing rate. An important part of the assessment is the analysis of prostate biopsies for Gleason classification, combining transrectal ultrasonography and intrarectal prostate palpation.

However, in the biochemical aspects of prostate cancer assessment, PSAs that are not alone or in any combination (eg,% fPSA, free PSA, complex PSA, etc.) are clinically relevant on an individual basis. It has been shown to be reliable enough to predict the pathological staging of existing prostate cancer. Therefore, biochemical markers that can add information about pathological staging on an individual basis would benefit from radical prostatectomy, i.e. have the best chance of being cured from prostate cancer. It will allow clinicians to make deaf patients more carefully selected.

[0022] Much attention has recently been paid to the detection of human glandular kallikrein 2 (hK2) in serum and prostate tissue samples as a potential novel marker. In our study, we used pathological staging 2a / b PCa (organ-localized cancer).
Was aimed at studying the ability of serum levels of hK2 to distinguish patients with extraprostatic extension (pathological staging ≧ 3a). One day prior to surgery, serum collection and rapid processing of samples without pre-manipulation of the prostate are performed under optimal analysis conditions, pathological testing of prostate removal specimens according to the Stanford protocol for accurate pathological staging. Offered.

The prostate cancer aggressiveness is reliably estimated and more accurate prior to any therapeutic decision to avoid overtreatment or undertreatment (eg, radical prostatectomy, radiation therapy or careful waiting). Simple tests are even needed to provide information. PSA has been established as an important prognostic parameter. However, not serum PSA value is not properly represent the pathological grading is in progress ^12. Because serum PSA production tends to decrease with increasing histological grading (dedifferentiation), even in the small extent of clinically localized disease, total PSA
⁶ was a poor prophet of the prognosis of prostate cancer. Only at highly elevated levels (eg, above 100 ng / ml) PSA clearly indicates the presence of advanced metastatic disease.

This is the first detailed clinical report demonstrating hK2 as a more suitable tumor marker to reflect dedifferentiation of prostate tumor cells. Of particular clinical significance is the ability of hK2 to identify poorly differentiated tumors over PSA in the mid-range of 3-15 ng / ml PSA. One of the main topics in treatment decisions made today, on the one hand, has a high risk of extracapsular disease (
To exclude tumors (particularly with seminal vesicle dilation) from local treatment with curative intent. On the other hand, overtreatment in the case of relatively non-aggressive diseases should be avoided, as data from the natural history of prostate cancer indicate.

Our results, shown below, are supported by the observations of Darson et al., Which show a more dramatic increase in hK2 immunohistochemical staining with increased histological staging compared to PSA. ²² . In addition, there are indications that RT-PCR of hK2 may predict final, positive lymph node status.

Experimental Topic Study I The purpose of the first study was to evaluate the use of hK2 or hK2 in combination with a form of PSA as a marker for staging PCa in patients.

In the first study, the preoperative serum concentration of hK2 was determined to be pathological staging 2a /
whether the patients with cancer b differ from those with pathological staging> 3a disease, and such as before surgery for organ-localized cancer for extra-prostatic extension of the tumor To study if it could help in predicting
We studied the serum levels of human glandular kallikrein 2 (hK2) in patients with prostate cancer treated with radical prostatectomy.

Materials and Methods Patient Selection and Evaluation Serum samples from 68 men scheduled for radical postpubic prostate removal for clinically localized prostate cancer one day prior to surgery Collected. None of the patients received hormonal treatment before surgery. Serum was collected before any manipulation of the prostate and transported to our lab, where it was stored at -80 C until analysis.

[0029] The prostate of histologic features: prostate, was prepared according to the Stanford protocol ^33. It was inked over the surface, fixed in formalin for at least 24 hours, and processed with a 3 mm step section technique. Gleason system uses the histologic grading ^34, and staging according to the second revision of the fourth edition of the TNM classification.

Detection of hK2: For the detection of hK2, we used a previously described three-step immunofluorescence assay. Briefly, primary antibodies that do not cross-react with hK2 are provided in excess to prevent free and total PSA from reacting in the next reaction step. Next, a secondary biotinylated antibody, which reacts only with hK2 because the corresponding epitope on PSA is blocked in the first step, is added and it is bound to streptavidin-coated microtiter wells. All PSA is removed by washing. In the third step, the europium-labeled antibody reacts with the immobilized hK2. Europium forms a fluorescent complex in proportion to the amount of hK2. Recombinant P
Immunological cross-reactivity measured by SA was less than 0.1%. The analytical detection limit defined by the 3 * SD inaccuracy of the zero calibrator is 0.01 ng / ml, while the functional detection defined as the level where the coefficient of variation (CV) between assays is less than 20% The limit was 0.03 ng / ml.

Detection of Total and Free PSA and% fPSA: To detect total and free PSA, we used DELFIA's ProS
The status Dual PSA-Total / free assay was used. The assay works with a sandwich-based technique. In the first stage, the total PS
A and free PSA bind equimolar to the solid anti-total PSA antibody. In the next step, the europium-labeled antibody binds to an antigenic site accessible only on the free PSA molecule. Both lanthanoids form fluorescent complexes that are proportional to the amount of free (europium only) and total (samarium) PSA. From both results, the total PS
The ratio of free PSA to A (% fPSA) is calculated.

Algorithms using hK2, total PSA and free PSA: For clinical analysis, we studied three algorithms aimed at combining hK2 with free PSA and total PSA: , HK2 concentration multiplied by total PSA and divided by free PSA.
And third, the concentration of hK2 divided by the concentration of free PSA. The analysis of the three algorithms is the first-[hK2] ^* [total PSA] /
[Free PSA]-was the most powerful algorithm in discriminating between organ- and non-organ-localized cancers, so we applied this algorithm in our subsequent studies and ignored the remaining two algorithms did.

Study Design and Statistical Summary of Data: Each analyte (hK2, total PSA, free PSA),% fPSA and [hK2] ^*
The average, range and standard error were calculated for the algorithm [total PSA] / [free PSA]. Means and ranges were compared in patients with organ-localized and non-organ-localized tumors. Calculations of the significance of the differences were performed using the Mann-Whitney U test. A p-value of 0.05 or less was considered significant. Once the results for hK2 and [hK2] ^* [total PSA] / [free PSA] were obtained, the results were compared to total PSA, free PSA and% fPSA. Square plot (FIG. 1a)
1c) represents the concentration of hK2, the algorithm of [hK2] ^* [total PSA] / [free PSA] and the total PSA of organ and organ non-localized cancer.

Of the 68 patients who underwent surgery, 38 patients had organ-localized cancer, and 30 men who had a close-up showed organ-non-localized cancer. Table 1 shows the average and p-values for the analytes and algorithms tested, which are hK2 alone and [hK2] ^*
The [total PSA] / [free PSA] algorithm gives the most accurate information in discriminating between organ- and non-organ-localized cancers, followed by total PSA, free PSA, and insignificant% fPSA That's a brief statement.

HK2 was 0.03 in 5 of 38 patients (= 13.1%) with organ-localized cancer and in 0 of 30 patients with non-organ-localized cancer.
No detectable at levels below ng / ml. The average hK2 concentration of all samples is 0.18
ng / ml (range: <0.03-0.94 ng / ml). In organ-localized cancers, the average hK2 concentration was 0.09 ng / ml, ranging from <0.03 to 0.67 ng / ml. In organ-non-localized cancer, the average hK2 concentration was 0.30 ng / ml (range 0.
04-0.94 ng / ml). The complete data sorted by pathological staging is shown in Table 2 and a box plot of hK2 concentration is shown in FIG. 1a. The Mann-Whitney U test showed a statistically highly significant difference in hK2 concentration between organ- and non-organ-localized cancers (p = 0.0001).

The average result of the algorithm of [hK2] ^* [total PSA] / [free PSA] is 1
. 54 (range: 0.06 to 10.16). In organ-localized cancers, the average value was 0.93 in the range of 0.06 to 5.79. For organ non-localized cancer, the average result was 2.31 (range: 0.30-10.16). Complete data, separated by pathological staging, is shown in Table 3, and a box plot of [hK2] ^* [total PSA] / [free PSA] results is shown in FIG. 1b. The Mann-Whitney U test showed a statistically highly significant difference between organ- and non-organ-localized cancers (p = 0.0005).

The total PSA concentration of all samples was 10.73 ng / ml (range: 3.34-62.3 ng / ml)
)Met. In organ-localized cancer, the average PSA concentration is between 3.34 and 24.1.
It was 7.5 ng / ml in the range of ng / ml. Average PSA in organ non-localized cancer
The concentration was 14.81 ng / ml (range: 3.43-62.3 ng / ml). The complete data sorted by pathological staging is shown in Table 4 and the box plot of total PSA results is shown in FIG. 1c. The Mann-Whitney U test showed a statistically significant difference between organ- and non-organ-localized cancers (p = 0.0023).
.

The results for free PSA and% fPSA are shown in Table 1. If we only use hK2, [hK
2] ^{* With the} better results revealed using the algorithm of [total PSA] / [free PSA] and total PSA, we created more detailed tables and box plots for free PSA and% fPSA. Stopped showing.

Our results on the detection of hK2 have led to several conclusions. At first,
The majority of patients with prostate cancer where hK2 is clinically localized (63/68)
Could be detected in On the other hand, the presence in five patients with serum levels below the limit of detection is evident not only in the evaluation of prostate cancer patients who were hK2 negative in our cohort but also those without prostate cancer. Need an assay with lower functional detection.
The second point is that patients with organ-localized cancer did not have undetectable hK2 levels.

An improved hK2 assay based on a monoclonal anti-PSA antibody was 0.1%
Had less than PSA cross-reactivity. That value is low enough to allow us to assess the clinical significance of specific measurement of hK2 in serum, where the median hK2 level is about 1.3-1.6% PSA concentration. Is equivalent to 0.05ng /
Despite the low functional sensitivity limit of ml (defined by a coefficient of variation of less than 20%), the assay was performed on the following subjects: all healthy controls, 5 with BPH
No immunoreactivity of hK2 was detected in 0% of men, 30% of patients with clinically localized PCa and 4% with clinically advanced PCa. Specifically, in order to properly assess subjects who do not have malignant prostate lesions,
0.04 ng / ml hK2 was added to all samples with undetectable hK2 immunoreactivity.
We have specified the level. This proved necessary to avoid introducing any inappropriate discrimination between men with BHP and cancer patients with hK2 levels of less than 0.05 ng / ml. The conclusion is that all undetected hK2
Supported by results obtained by using a value of 0 at the level or by excluding patients with hK2 concentrations below the limit of detection (results not shown).

In our study, the inclusion of hK2 in the diagnostic preoperative workup of radical prostatectomy patients improved the classification of organ-localized and non-organ-localized cancers. Either hK2 alone or the algorithm [hK2] ^* [total PSA] / [free PSA] was compared to total PSA, free PSA and the relationship of free PSA to total PSA (% fPSA) for this purpose. There was a statistical advantage for
Thus, the inclusion of hK2 in the preoperative biochemical assessment of prostate cancer would be a useful tool for improved selection of patients with histologically proven prostate cancer.

Study II The second study objective was to evaluate the use of the hK2 measurement itself for the grading of PCa in patients. Materials and Methods The study population consisted of 122 consecutive patients with histologically proven prostate cancer. Histology-based diagnosis was made on tissue from a six-section biopsy guided by transrectal ultrasound of the prostate and / or all glands obtained after radical prostatectomy. The malignant grades are well differentiated (G1, n = 35), moderately differentiated (G2
, N = 61) or poorly differentiated (G3, n = 26) carcinoma. The patient,
He had never received antiandrogen treatment, transurethral resection, radical prostatectomy or radiation therapy.

Blood samples were obtained prior to any prostate procedures. After clot formation, the samples were centrifuged, and serum was collected and frozen at -70 ° C. The sample was melted immediately before the measurement. hK2 measurements are already described ^41, and were performed by indirect immunofluorescence assay based on indirect PSA capture step. Analytical sensitivity (background + 3SD) is 0.01 ng / ml and functional sensitivity is 0.05
ng / ml (defined as the intra-assay coefficient of variation of 20% or less) (25 μl
Aliquots of serum) or 0.02-0.03 ng / ml (50 μl
If an aliquot of serum was used). The cross-reactivity of this assay with PSA reached less than 0.1% by using two capture antibodies to prevent PSA from being plugged in this assay.

[0044] Total PSA and free PSA were obtained from commercially available Delita ProS for immunofluorescence measurement.
status PSA Free / Total Kit (Wallac Oy, Tu
rku, was determined by Firland) ^42.

The alpha-1-antichymotrypsin bound to PSA (PSA ACT) also measures to those described previously by similar immunofluorescence assay ^43, however, anti-PS
A monoclonal IgG (H117) was used as a capture antibody and Eu-labeled anti-A
CT monoclonal IgG (241) was used as a detection antibody.

Statistical analysis: A multivariate logical regression analysis was performed to detect the best conditions for the tumor markers. Statistical analysis was performed using commercially available computer software. In each tumor discrimination group, the median and average (± SD) of total PSA, free PSA, PSA-ACT and hK2 were calculated. The same procedure was performed for the following combinations: free / total PSA; hK2 / free PSA; (hK2 / free PSA) x (total PSA / free PSA); free PSA / PSA-ACT; PSA-ACT. / Total PSA; hK2 / total and free PSA / (total PSA x hK2). we,
To determine the statistical significance of differences between populations, a nonparametric man-
The Whitney U test was used.

For all analytes, a p-value less than 0.05 was considered statistically significant.

Results of Study II The descriptive statistics (median, mean ± SD) of different markers and their combinations in G1, G2 and G3 tumors are shown in Table 5. Total PSA was approximately 2-fold (13.1 to 2) from G1 to G2 tumors (p = 0.0002) and from G2 to G3 tumors.
6.5 ng / ml). However, the latter increase was not significant (p = 0.
18). In contrast, hK2 also increased from G2 to G3 by a factor of 3 (p = 0.0
2). The ratio of free PSA to total PSA was reduced in G1 compared to G2 (0.15 to 0.10, p = 0.007). The statistically significant difference is G2
It was not found among the G3 population (0.10 vs. 0.11, p = 0.93). However, the ratio of hK2 / free PSA was also distinguished between G2 and G3 tumors (0.1
1: 0.22, p = 0.002). In the multivariate regression analysis, hK2 ((hK2
Combinations containing (/ free) x (total / free); hK2 / total; free / (total x hK2)) were also distinguished between G2 and G3 in a statistically significant manner. The results from G1 versus G3 are shown in Table 5.

Table 6 compares the combination of well differentiated and moderately differentiated prostate cancer (G1 + G2) with poorly differentiated tumor (G3) by median. The statistically most significant differences between these populations were hK2 (p = 0.001), hK2 / free PSA (p = 0.0003)
And free PSA / (total x hK2) (p = 0.0004). A p-value of 0.01 for total PSA was recorded, while free P
The SA ratio failed to discriminate between the two groups.

From a clinical point of view, cancers with a total PSA range of 3-15 ng / ml form the most important group. In this range, the differentiated carcinomas had total PSA (10.6 vs. 7.8 ng / ml).
, P = 0.20), free PSA (1.19 vs. 0.85 ng / ml, p = 0.55)
Or G1 / G by PSA-ACT (9.3 vs. 7.3 ng / ml, p = 0.22).
Two tumors could not be distinguished (Table 7). In contrast, hK2 is poorly differentiated G
In three tumors, there was a 2.9-fold increase from 0.08 ng / ml (G1 / G2) to 0.23 ng / ml (p = 0.03). Furthermore, the ratio of hK2 / free PSA is G1 / G2 and G3
(0.09 vs. 0.17, p = 0.02), but free PS
A /% of total PSA failed (0.12 vs 0.10.p = 0.3).

The above results show that hK2 significantly improved the identification of more aggressive (G2 to G3) tumors compared to total PSA, free PSA and PSA-ACT. It is important to note that improved detection of aggression was also seen in the mid-range of total PSA (3-15 ng / ml). Thus, hK2 itself is a useful tool for pre-treatment decision analysis.

It will be appreciated that the methods of the present invention may be embodied in various forms of embodiment, a few of which are disclosed herein. It will be apparent to one skilled in the art that other embodiments exist and do not depart from the spirit of the invention. Accordingly, the described embodiments are illustrative and should not be regarded as limiting.

[Table 1]

[Table 2]

[Table 3]

[Table 4]

[Table 5]

[Table 6]

[Table 7]

[Table 8]

[Table 9]

[Table 10]

[Table 11]

[Table 12]

[Table 13]

[Table 14]

[Brief description of the drawings]

FIG. 1a shows the concentration of hK2.

FIG. 1b shows the algorithm of [hK2] ^* [total PSA] / [free PSA].

FIG. 1c shows the total PSA of organ-localized (oc) and organ-non-localized (noc) cancers.

[Procedural Amendment] Submission of translation of Article 34 Amendment

[Submission date] May 17, 2001 (2001.5.17)

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] Claims

[Correction method] Change

[Contents of correction]

[Claims]

────────────────────────────────────────────────── ─── Continuing on the front page (72) Inventor Wrecker, Franz Helmut Rihardt, Switzerland, Zeher-5016 Oberer Rinsbach, Erzbergbeek 14 (72) Inventor Kubiatokowski, Makiei Kurziztov Poland, PI-80-809 Guda Nsk, Ulica Bitosa 21/49

Claims (9)

[Claims] 1. A method for staging prostate cancer, i.e., for distinguishing a patient's organ-localized prostate cancer (PCa) from non-organ-localized PCa, comprising the step of treating a patient's human glandular kallikrein 2 ( hK2) and optionally also a prostate specific antigen (P
The method wherein the fluid concentration of SA) is determined and hK2 is used as a marker to distinguish patients with organ-localized PCa from patients with organ-non-localized PCa. 2. The method according to claim 1, wherein hK2 is used alone as said marker. 3. The method according to claim 1, wherein a combination of hK2 and PSA (where PSA means free PSA, complex PSA or total PSA) is used as the marker. Method. 4. The method according to claim 3, wherein the marker is an algorithm of hK2 × total PSA / free PSA. 5. A method for grading prostate cancer, that is, for distinguishing a patient having aggressively progressing prostate cancer from a patient having a less aggressively progressing prostate cancer. And the patient's human glandular kallikrein 2
A method characterized in that the body fluid concentration of (hK2) is determined and hK2 is used alone as said marker. 6. The method of claim 5, wherein the body fluid concentration of the patient's prostate specific antigen (PSA) is also determined and the patient has a total PSA in the range of 1-20 ng / ml. 7. The method of claim 6, wherein the patient has a total PSA in the range of 3-15 ng / ml. 8. The method according to claim 5, 6 or 7, characterized in that on the one hand the patients with well-differentiated and moderately differentiated PCa and on the other hand the patients with poorly differentiated PCa are distinguished. 9. The method according to claim 5, 6 or 7, characterized by distinguishing between patients having, on the one hand, moderately differentiated PCa and, on the other hand, poorly differentiated PCa.