Materials and methods

Study design and sample size

This was a hospital-based cross-sectional observational study including 25 patients per group (i.e., a total sample size of 50, assuming equal group sizes of prostate adenocarcinoma and BPH cases).

Ethical approval

Ethical approval for the study was obtained from the Shri BM Patil Medical College, Hospital and Research Centre, Vijayapura Institutional Ethics Committee (reference number: BLDE(DU)IEC-SBMPMC/124/2023-24).

Inclusion and exclusion criteria

Histologically diagnosed cases of prostate adenocarcinoma and BPH were included in the study, and metastatic carcinomas of the prostate were excluded.

Data collection

Specimens of the prostate, such as prostatic needle biopsy (10 samples) and TURP CHIPS (chips of transurethral resection of the prostate, 15 samples), were received in the Histopathology Section, Department of Pathology, BLDE (Deemed to be University), Shri BM Patil Medical College, Hospital and Research Centre, Vijayapura, Karnataka. Information regarding Clinical details, PSA levels, and findings on imaging, if any, was obtained. The tissue was preserved in 10% buffered formalin and processed routinely. Three micro-thick sections were prepared from each tissue block. One section was stained with hematoxylin and eosin (H&E) for morphologic diagnosis, and Gleason’s score and grade, and the other two sections were mounted on poly-L-lysine-coated slides, which were used for p53 and Ki-67 immunohistochemical staining.

Immunohistochemical analysis

Positive and negative controls were processed at the same time. Strong brown staining in cell nuclei indicated positive results. Staining was quantified by the percentage of tumor cells reacting to the antibody. Slides were first scanned at ×40 magnification to locate regions with the highest positive cell density. These areas were then assessed at ×400 magnification, where the proportion of positive cells to total cells was determined. At least 500 cells were counted per slide, including only those positive for the target marker.

Statistical analysis

The data obtained were entered into a Microsoft Excel (Redmond, Washington, USA) sheet, and statistical analysis was performed using the Statistical Package for Social Sciences (version 20). Results are presented as mean ± standard deviation (SD), counts and percentages, and diagrams. Normally distributed continuous variables between two groups were compared using an independent t-test, and the Mann-Whitney U test was used for not normally distributed variables. Categorical variables between two groups were compared using a chi-square test. A p<0.05 was considered statistically significant. All statistical tests were two-tailed.