Study on some histopathological features and expression of some immunohistochemical markers in prostatic carcinoma

Objectives: To determine some histopathological features and evaluate the expression of some IHC markers in prostate carcinoma (Pca). Subjects and methods: Histopathological study was performed on 84 specimens by using hematoxylin and eosin (H.E) stained tissue sections. IHC study was performed on 33 specimens by using IHC stained tissue sections with PSA, CK34βE12, p63, CK7, CK5/6, actin monoclonal antibodies.

Journal of military pharmaco-medicine n08-2017 STUDY ON SOME HISTOPATHOLOGICAL FEATURES AND EXPRESSION OF SOME IMMUNOHISTOCHEMICAL MARKERS IN PROSTATIC CARCINOMA Vi Thuat Thang*; Tran Ngoc Dung*; Nguyen Dinh Tao** Nguyen Ngoc Hung*; Tran Ngoc Anh** SUMMARY Objectives: To determine some histopathological features and evaluate the expression of some IHC markers in prostate carcinoma (Pca) Subjects and methods: Histopathological study was performed on 84 specimens by using hematoxylin and eosin (H.E) stained tissue sections IHC study was performed on 33 specimens by using IHC stained tissue sections with PSA, CK34βE12, p63, CK7, CK5/6, actin monoclonal antibodies Results: Prostatic adenocarcinoma accounted for 96.4% (acinar type was the most common) and urothelial carcinoma of the prostate accounted for 3.6% Gleason scores - were the most common Of the 84 patients, 60 (73.2%) had high grade prostatic intraepithelial neoplasia (HGPIN) associated adenocarcinoma There were 39% of adenocarcinoma with perineural invasion and 53.7% of adenocarcinoma with mucinous secretions IHC showed 31 cases (94%) of Pca which were positive for PSA and cases (6%) of urothelial carcinoma of the prostate were negative for PSA The lower Gleason grade was stronger positive for PSA was and vice versa All the tumor cells were negative for 34βE12, p63, CK7, CK5/6 and actin Conclusion: Our data indicate that most of Pca are prostatic adenocarcinoma and Gleason scores - are the most common scores HGPIN is strongly associated with adenocarcinoma Other features are commonly seen in prostatic adenocarcinoma which are blue-tinged mucinous secretions and perineural invasion IHC shows all of prostatic adenocarcinomas are positive for PSA, urothelial carcinoma of the prostate is negative for PSA The staining intensity of the tumor cells for PSA is stronger in the lower grade adenocarcinomas and vice versa * Keywords: Adenocarcinoma of the prostate; Immunohistochemistry INTRODUCTION Prostatic carcinoma is one of the malignant tumours and often found in men over the age of 65 Nowadays, this kind of tumour has the tendency to increase dramatically in almost every country around the world Histopathologically, adenocarcinomas of the prostate range from well-differentiated gland forming cancers, where it is often difficult to distinguish them from benign prostatic glands, to poorly differentiate tumours, difficult to identify as being of prostatic origin [8] Currently in about two-thirds of these difficult cases, an appropriate diagnosis can be made due to IHC stains with the use of antibodies [4] In Vietnam, the studies on histopathological features and the expression of IHC markers in carcinoma of the prostate are not adequate to the demand [1] The aim of this study is: To determine some histopathological features and evaluate the expression of some IHC markers in Pca * 103 Military Hospital ** Vietnam Military Medical University Corresponding author: Vi Thuat Thang (vithuatthang@yahoo.com) Date received: 23/03/2017 Date accepted: 27/09/2017 222 Journal of military pharmaco-medicine n08-2017 SUBJECTS AND METHODS Subjects Eighty four consecutive prostate glands operated by using transurethral resection of the prostate (TURP) at Urology of 103 Military Hospital from June, 2008 to July, 2017 Methods Specimens were fixed in 10% neutral buffered formalin, embeded in paraffin and cut on the microtoma Tissue sections were stained with H.E and IHC stained with the use of PSA, CK5/6, CK34βE12, p63, CK7, actin monoclonal antibodies The results were presented and discussed Tissue sections of 33 cases were IHC stained with use of PSA, CK5/6, CK34βE12, p63 and actin monoclonal antibodies, in which there were 31 cases of adenocarcinoma and cases of urothelial carcinoma of the prostate - Determine the ratio of prostatic carcinomas which were positive for PSA, CK34βE12, p63, CK7, CK5/6 and actin - Distribute staining intensity of tumor cells for PSA according to Gleason grade RESULTS AND DISCUSSION Distribution of patients’ age Table 1: Distributed results of Pca among age group (n = 84) Age group No of patients (n = 84) Ratio (%) 40 - 49 1.19 50 - 59 4.76 60 - 69 20 23.81 70 - 79 36 42.86 ≥ 80 23 27.38 Total 84 100% * Distribution the age: Ages of patients were divided into groups with the gap of 10 years (< 50, 50 - 59, 60 - 69, 70 - 79 and ≥ 80) * Study on histopathology: - Determine the types of histopathology of the Pca according to the 2004 WHO histological classification of tumours of the prostate - Evaluate the association between adenocarcinoma and HGPIN - Grade the adenocarcinomas according to the Gleason grading system - Group Gleason scores into diffentiation categories - Determine some histopathological features of prostatic adenocarcinoma * Study on the expression of some IHC markers in prostatic carcinoma: This table showed the incidence of Pca increased with age group The 70 - 79 age group was the highest with 42.86%, the 40 - 49 age group was the lowest with 1.19% These results showed that prostatic carcinoma was rarely found in men below the age of 50 [1, 6, 8] Our results of distributed Pca among age group are the same as the outcome of study carried out by Nguyen Van Hung [1], Nguyen Buu Trieu [2] and others [7] 223 Journal of military pharmaco-medicine n08-2017 Study on histopathology Table 2: Determination of types of histopathology of the Pca according to the 2004 WHO histological classification (n = 84) Types of histopathology No of patients Ratio (%) Adenocarcinoma Acinar adenocarcinoma Dductal adenocarcinoma Urothelial carcinoma Squamous carcinoma Basal cell carcinoma 82 80 2 0 96.4 97.6 2.4 3.6 Total 84 100% This table showed adenocarcinoma accounted for 96.4% (in which acinar adenocarcinoma was 97.6%, ductal adenocarcinoma was 2.4%) and urothelial carcinoma of the prostate accounted for 3.6% These results were the same as the results found by Nguyen Van Hung [1] and others Table 3: Association between adenocarcinoma and HGPIN (n = 82) Histopathology Adenocarcinoma is associated with HGPIN Adenocarcinoma is not associated with HGPIN Total No of patients Ratio (%) 60 22 73.2 26.8 82 100 % Adenocarcinoma associated with HGPIN and accounted for 73.2%, much higher than adenocarcinoma, did not associate with HGPIN (26.8%) Nguyen Van Hung [1] also found that 76.2% of prostatic carcinoma contained HGPIN Clinical significance of HGPIN showed strong association with Pca [5] 224 HGPIN was a predictive factor which was valuable in the diagnosis of adenocarcinoma of the prostate, especially HGPIN with adjacent atypical glands (PINATYP) [8] HGPIN in needle biopsy tissue was a risky factor for the subsequent carcinoma detection after re-biopsy [1, 2, 5, 9] Table 3: Grouping Gleason score into diffentiation categories (n = 82) Differentiated grade Gleason score No of patients Ratio (%) Well differentiated Moderately differentiated Poorly differentiated 2-4 5-7 14 58 17.1 70.7 - 10 10 12.2 82 100% Total Two cases of urothelial carcinoma of the prostate were not included in this scoring system Gleason score tumors - were the highest ratio (70.7%), Gleason score tumors - and - 10 were 17.1% and 12.2%, respectively These results were the same as the study carried out by Nguyen Van Hung [1], Bostwick, Oesterling, Zincke [1, 3, 5, 10] Table 5: Distributed ratio of adenocarcinomas according to malignant specific features (n = 82) Malignant specific features Number Ratio (%) Perineural invasion 32 39% Mucinous fibroplasia 11% Glorumerulation 10 12.2% or of malignant specific features 7.3% No malignant specific features 25 30.5% 82 100% Total Journal of military pharmaco-medicine n08-2017 The table showed adenocarcinoma with perineural invasion was found in 39% of cases, adenocarcinoma with glorumeration and mucinous fibroplasia were found in 12.2% and 11%, respectively Adenocarcinoma with or malignant specific features were found in 7.3% of cases while adenocarcinoma with no malignant specific features were found in 30.5% of cases There were only features that are in and of themselves diagnostic of cancer These were perineural invasion, mucinous fibroplasia and glorumeration Perineural invasion specimens by prostate cancers were seen in radical prostatectomy specimens in 75 - 84% of cases Most studies indicate that its pesesence corellated with extraprostatic extension (38 - 93%) Recent data suggest that this finding may indipendently predict lymph-node metastasis and prognosis after surgery [8] Table 6: Distributed ratio of adenocarcinoma according to intraluminal features (n = 82) Intraluminal features Number Ratio (%) Crystalloids 9.7% Blue-tinged mucinous secretions 44 53.7% Crystalloids and mucinous secretions 18 22% No crystalloids and mucinous secretions 12 14.6% 82 100% Total Crystalloids were found in 9.7% of cases, blue-tinged mucinous secretions (53.7%), crystalloids and mucinous secretions (22%), no crystalloids and mucinous secretions (14.6%) Crystalloids, despite not diagnostic of carcinoma, were more frequently found in low grade cancer than in benign glands [11] Young [9] also showed that the incidence of crystalloids in the adenocarcinoma ranged 10 - 62% Blue-tinged mucinous secretions seen in H.E stained sections were additional finding seen preferentially in cancer, especially low grade cancer [8] Fig.1: Well- differentiated adenocarcinoma, Gleason grade (Arrow: mucinous secretions) (No S2835 x 200.HE) Fig.2: Poorly differentiated adenocarcinoma, Gleason grade (Arrow: perineural invasion) (No.S3475 x 400.HE) 225 Journal of military pharmaco-medicine n08-2017 Study on immunohistochemistry Table 7: Ratio of Pca which were positive for PSA, 34βE12, p63, CK7, CK5/6 and actin (n = 33) Type of marker Number of positive cases Ratio (%） PSA 31/33 94% 34βE12 0/33 0% p63 0/33 0% CK7 0/33 0% CK5/6 0/33 0% Actin 0/33 0% The study results were consistent with the findings of other studies [1, 4, 8] Table 8: Staining intensity distribution of tumor cells with PSA according to Gleason grade (n = 31) Intensity Number (1+) (2+) 15 (3+) (4+) Tổng 31 Grade Grade Grade Grade 5 15 9 15 (Positive: +; Faint: 1+; Moderate: 2+; Strong: 3+; Very strong: 4+) IHC showed tumor cells which were strong positive for PSA in the areas of Gleason grade 2, moderate positive for PSA in the areas of Gleason grade 3, faint positive for PSA in the areas of Gleason grade and The study results were consistent with the findings of other studies The finding is that the more the differentiated tumor cell is, the stronger the positive intensity for PSA is and vice versa [1, 4, 8] Fig 3: Gleason grade (Arrow: strong positive) No N3351 x200 PSA 226 Fig 4: Gleason grade (Arrow: moderate positive) No N3351 x200 PSA Fig 5: Gleason grade (Arrow: basal cells (2+) for 34βE12 in benign glands) (No N2519 x200 34βE12) Journal of military pharmaco-medicine n08-2017 CONCLUSION * Histopathological features: Prostate carcinoma is common in the older men Most of Pca are adenocarcinoma in which acinar adenocarcinoma type is the most common, ductal adenocacinoma is a rare variant of prostatic adenocarcinoma Gleason scores - are the most common Adenocarcinoma is strongly associated with HGPIN Other features commonly seen in prostatic adenocarcinoma which are blue-tinged mucinous secretion and perineural invasion * Expression of some immunohistochemical markers: IHC shows all of prostatic adenocarcinomas are positive for PSA, urothelial carcinomas of the prostate are negative for PSA The staining intensity of the tumor cells for PSA are stronger in the lower grade adenocarcinomas and vice versa Tumor cells are negative for CK34βE12, p63, CK7, CK5/6 and actin REFERENCES Nguyễn Văn Hưng Nghiên cứu mô bệnh học q sản lành tính, tân sản nội biểu mơ ung thư biểu mô tuyến tiền liệt Luận án Tiến sỹ Y học 2005 Nguyễn Bửu Triều, Nguyễn Kỳ, Trần Đức Thọ Các biểu lâm sàng ung thư tuyến tiền liệt giai đoạn muộn Chẩn đốn điều trị Tạp chí Ngoại khoa 1991, 1, tr.9-13 Bostwick D.G Prostatic intraepithelial neoplasia Urol (suppl) 1999, 34 (6), pp.23-27 Buchwalow I.B, W Böcker Immunohistochemistry: basics and methods Springer Science & Business Media 2010 Oesterling J.E, Brendler C.B, Epstein J.I et al Correlation of clinical stage, serum prostatic acid phosphatase and preoperative Gleason grade with final pathological stage in 275 patients with clinically localized prostate cancer J Urol 1987, 138, pp.92-96 Stanley L.Rbbins M.D Pathologic basis of disease W.B Saunders Company 1974, pp.1190-1198 Tannenbaurn M, Galang C.F, Park C.H et al Dysplasia of the prostate, a precursor lesion of prostatic cancer Ann Urol 1991, pp.27-49 John N Eble, Guido Sauter, Jonathan I Estain, Isabell A Sesterhenn Pathology and genetics of tumours of the urinary system and male genital organs IARCPress Lyon 2004, chapter 3, pp.159-215 Young R.H, Srigley J.R, Amin M.B, Ulbright T.M, Cubillia A.L Tumors of the prostate gland, seminal vesicles, male urethra, and penis 3rd, Fascicle 28 Raven 2000, pp.1-344 10 Zincke H, Farrow G.M, Myers R.P et al Relationship between grade and stage of adenocarcinoma of the prostate and regional pelvic lymph node metastases J Urol 1982, 128, pp.498-501 227 ... categories - Determine some histopathological features of prostatic adenocarcinoma * Study on the expression of some IHC markers in prostatic carcinoma: This table showed the incidence of Pca increased... common Adenocarcinoma is strongly associated with HGPIN Other features commonly seen in prostatic adenocarcinoma which are blue-tinged mucinous secretion and perineural invasion * Expression of some. .. is common in the older men Most of Pca are adenocarcinoma in which acinar adenocarcinoma type is the most common, ductal adenocacinoma is a rare variant of prostatic adenocarcinoma Gleason scores