Page 43 - ARNM-3-3

Advances in Radiotherapy
& Nuclear Medicine PI3K and Akt in androgen-independent PCa

In 1941, Huggins and Hodges reported that castration The exclusion criteria were as follows:
2
can delay the progression of PCa and alleviate patients’ (v) Patients without a needle biopsy
symptoms. To date, androgen deprivation therapy (ADT) (vi) Patients with a history of postoperative radiotherapy
remains the primary treatment for advanced disease. or endocrine therapy
However, over time, tumors inevitably progress into a (vii) Patients with chronic or acute inflammation
castration-resistant stage, in which androgen is no longer (viii) Patients diagnosed with other malignant tumors
effective in suppressing PCa progression. (ix) Patients with hematological or autoimmune diseases
The accelerated proliferation and differentiation (x) Patients with incomplete original clinical data.
of cancer cells reduce their dependence on androgen 2.2. Immunohistochemical analysis
signaling, ultimately leading to disease progression or
metastasis through mechanisms that are not yet fully The expression levels of AR, PI3K, and Akt were
™
understood. Although the mechanisms underlying analyzed immunohistochemically using the MaxVision
treatment failure remain unclear, the phosphatidylinositol- method. Paraffin sections were dewaxed with xylene,
3-kinase (PI3K)/protein kinase B (Akt) signaling pathway dehydrated through a graded alcohol series to water,
has been implicated in the onset and progression of many and then incubated with 3% hydrogen peroxide to
tumors. The observed correlations among the androgen eliminate endogenous peroxidase activity. Non-specific
receptor (AR), PI3K, and Akt may aid in identifying binding sites were blocked using normal goat serum. The
molecular markers that differentiate androgen-dependent sections were incubated with rabbit anti-human primary
PCa (ADPC) from androgen-independent PCa (AIPC). antibodies (diluted 1:200) (Abcam, United States) at
Furthermore, these insights may facilitate the discovery room temperature for 2 h, followed by incubation with
™
of new therapeutic targets for the treatment of refractory the MaxVision polymer detection reagent (Abcam,
AIPC. United States) at room temperature for 30 min. The
reaction was terminated by rinsing with distilled water
2. Methodology after chromogenic development. Subsequently, cover slips
were placed on the sections for microscopic observation
2.1. Patients and imaging. For negative controls, adjacent sections were
From January 2012 to December 2017, paraffin-embedded processed in the same manner, with either normal goat
tissue specimens or needle biopsies of PCa were serum or phosphate-buffered saline replacing the primary
retrospectively collected at the First Affiliated Hospital antibody. The sections were observed and recorded using
of Xi’an Jiaotong University, and the corresponding an Olympus BX51 microscope (Olympus, Japan).
pathological sections were reviewed. Classification and
postoperative clinicopathological staging were conducted 2.3. Determination of immunohistochemical results
according to the 2014 Prostate Gland Gleason classification Positive expressions of AR, PI3K, and phosphorylated Akt
standard. A total of 76 cases were included: 38 in the AIPC were indicated by distinct staining patterns: AR localization
3
group and 38 in the ADPC group. This study was approved to the nucleus (brownish-yellow), PI3K to the cytoplasm
by the Medical Ethics Review Board of the Affiliated (light yellow or brownish-yellow), and phosphorylated Akt
Hospital of Xi’an Jiaotong University, and all patients to both the cytoplasm and nucleus (brownish-yellow).
provided informed consent to participate in the study. For the immunohistochemical scoring standard, four

The inclusion criteria were as follows: different fields in each section were randomly selected and
(i) Serum testosterone is at the castration level; serum scanned under a 40× objective lens. In each field, 200 cancer
prostate-specific antigen (PSA) levels measured for cells were assessed. The percentage of positively stained cells
3 consecutive times (with an interval of at least 2 weeks) was calculated using the following formula (Equation I):
are higher than the reference value (0.2 ng/dL) and Number of positive cells
exceed 50% of the lowest value Percentage of positive cells = (I)
(ii) The concentration of serum testosterone shows a 200 cells
decreasing trend
(iii) After discontinuing antiandrogen therapy or switching Staining evaluation was independently performed by
to second-line antiandrogen drugs, serum PSA levels two authors. The proportion of positive cells was scored on
do not decrease and even increase instead a scale from 0 to 4 as follows: 0 for negative, 1 for ≤10%, 2
(iv) After at least 4 weeks of antiandrogen withdrawal for 11 – 50%, 3 for 51 – 75%, and 4 for >75%.
therapy, progression of bone or soft tissue metastases Staining intensity was scored on a scale from 0 to 3 as
is observed. follows: 0 for “no staining,” 1 for “light yellow and weak

Volume 3 Issue 3 (2025) 35 doi: 10.36922/ARNM025160018