Development of an algorithm for biomedical image analysis of the spatial organization of collagen in breast cancer tissue of patients with different clinical status.

Collagen, the main component of the tumor microenvironment, plays a key role in the development of breast cancer (BCa); however, the specific changes in its spatial organization during tumor progression have not been definitively elucidated. The existing and available methods for assessing the morphometric parameters of the stroma's fibrous component are insufficient for a detailed description of the state of collagen fibers and for assessing its changes to evaluate the aggressiveness of the BCa course. The aim of the work was to develop an algorithm for microphoto analysis to assess the spatial organization of collagen in BCa tissue of patients with different clinical statuses. The study was conducted on 60 tissue samples of stage I-II BCa. The processed images were analyzed using the software packages CurveAlign v4.0 and imagej. We established that the increase in BCa stage and the decrease in tumor differentiation grade are associated with decreased length, width, and straightness of collagen fibers, as well as their increased density. The formation of an aggressive basal molecular BCa subtype was accompanied by an increase in tumor-stroma ratio. The obtained results indicate the possibility of practical application of the developed algorithm for evaluating the spatial organization of collagen in BCa tissue to predict the aggressiveness of the disease course.

Osteonectin (SPARC) prognostic value in prostate cancer.

BACKGROUND: Prostate cancer (PCa) is common malignancy among men worldwide. To date only few molecular markers are available to predict its course and outcome. SPARC is considered to be promising prognostic marker of PCa due to its involvement in various cancer processes. MATERIALS AND METHODS: study was conducted on PCa surgical primary tumor samples, obtained from 84 patients. Level of SPARC mRNA expression was estimated using RT-qPCR. To identify SPARC protein (osteonectin) in prostate tissue, immunohistochemical analysis was conducted. Bioinformatical analysis was performed on UALCAN and TNMplot resources. RESULTS: bioinformatical analysis demonstrated that SPARC mRNA levels are decreased in PCa samples, in comparison to normal tissue. In patients with lymph node metastases its levels are 1.26 times higher; p = 4.66E-02, than in N0 category. Ex vivo study demonstrated that SPARC expression was elevated on both mRNA and protein levels in PCa patients with lymph node metastases (by 2.34 and 1.91, respectively, p < 0.05). We established higher levels of SPARC mRNA and protein in PCa patients with T3 tumors, as well as high Gleason score. Estimation of survival rates demonstrated that PCa patients with a high level of SPARC mRNA and protein have decreased overall 2-year survival. CONCLUSIONS: SPARC protein was overexpressed on mRNA and protein levels in patients with presence of lymph node metastases and higher Gleason score of tumors. Also, both mRNA and protein upregulation were associated with worse survival rates. The current study has therefore provided further evidence that SPARC is indeed linked to the prognosis and aggressiveness of human PCa.

Combined Dextran-Graft-Polyacrylamide/Zinc Oxide Nanocarrier for Effective Anticancer Therapy in vitro.

Introduction: Cancer chemotherapy faces two major challenges - high toxicity of active substances and tumor resistance to drugs. Low toxic nanocarriers in combination with anticancer agents can significantly increase the effectiveness of therapy. Modern advances in nanotechnology make it easy to create materials with the necessary physical and chemical properties. Methods: Two hybrid nanosystems of dextran-polyacrylamide/ zinc oxide nanoparticles (D-PAA/ZnO NPs) were synthesized in aqueous solution with zinc sulphate (D-PAA/ZnO NPs (SO42-)) and zinc acetate (D-PAA/ZnO NPs (-OAc)). The light absorption, fluorescence, dynamic light scattering and transmission electron microscopy for nanocomposite characterization were used. MTT, neutral red uptake and scratch assays were selected as fibroblasts cytotoxicity assays. Cytotoxicity was tested in vitro for normal fibroblasts, MAEC, prostate (LNCaP, PC-3, DU-145) and breast (MDA-MB-231, MCF-7) cancer cells lines. Immunocytochemical methods were used for detection of Ki-67, p53, Bcl-2, Bax, e-cadherin, N-cadherin and CD44 expression. Acridine orange was used to detect morphological changes in cells. Results: The radius of ZnO NPs (SO42-) was 1.5 nm and ZnO NPs (-OAc) was 2 nm. The nanosystems were low-toxic to fibroblasts, MAEC. Cells in the last stages of apoptosis with the formation of apoptotic bodies were detected for all investigated cancer cell lines. Proapoptotic proteins expression in cancer cells indicates an apoptotic death. Increased expression of E-cadherin and N-cadherin was registered for cancer cells line LNCaP, PC-3, DU-145 and MCF-7 after 48 h incubation with D-PAA/ZnO NPs (SO42-). Conclusion: The nanosystems were low-toxic to fibroblasts, MAEC. The D-PAA/ZnO NPs nanosystem synthesized using zinc sulphate demonstrates high cytotoxicity due to destruction of various types of cancer cells in vitro and potentially increases adhesion between cells. Thus, our findings indicate the selective cytotoxicity of D-PAA/ZnO NPs against cancer cells and can be potentially used for cancer treatment.

Benign and malignant prostate neoplasms show different spatial organization of collagen.

AIM: To compare the indicators of the spatial organization of collagen and its regulating factors between benign and malignant prostate neoplasms. METHODS: The study involved tumor tissue samples from 40 patients with stage II-III prostate cancer (PCa) and 20 patients with benign prostatic hyperplasia (BPH). The localization of collagen was determined with a Masson trichrome stain. To establish quantitative indicators of the spatial organization of collagen, morphometric studies were carried out with the CurveAlign and ImageJ programs. RESULTS: PCa tissue had two times lower collagen density (P<0.0001) and 1.3 times lower levels of collagen alignment (P=0.018) compared with BPH tissue. In PCa tissue, collagen fibers were shorter (by 24.2%; P<0.001) and thicker (by 15.5%; P<0.001). PCa tissue samples showed significantly higher levels of metalloproteinase (MMP)-2 (by 2.4 times; P=0.001), MMP-8 (by 2.3 times; P=0.007), and MMP-13 (by 1.9 times; P=0.004). CONCLUSIONS: Collagen matrix spatial organization features, as well as its regulatory factors, could be potential biomarkers of malignant prostate neoplasms.

Mast cells as a tumor microenvironment factor associated with the aggressiveness of prostate cancer.

We aimed to investigate the relationship between the degree of mast cells' (MCs) infiltration and clinicopathological features of prostate cancer (PCa) malignancy and to find out the possible mechanisms of the involvement of these cells in the formation of the aggressive course of the PCa development. The study was conducted on the clinical material of 60 patients with PCa of stages II-III. MCs in the PCa tissue were determined by a histochemical method using toluidine blue. The expression of osteopontin (OPN) was studied by the immunohistochemical method. The expression of miRNA-21, -126, -146a, -181a, and -221 was investigated by quantitative real-time PCR. Statistical processing of the results was performed using the GraphPad Prism 8 program. Our results demonstrated that the increased level of infiltration and degranulation of MCs in the PCa tissue was associated with such indices of the malignancy of the tumor process as the Gleason score and the preoperative PSA level in the blood serum of patients. A high level of MCs infiltration of the PCa tissue was associated with a significant decrease in the two-year recurrence-free survival rates of the patients by 23.3% (р=0.0455). A high degree of MCs infiltration of the PCa tissue was associated with 1.2 times (p=0.0347) higher level of OPN expression and 1.7 (p=0.0051) and 1.65 (p=0.0087) times lower levels of miR-126 and miR-181a expression, respectively. The obtained results indicate the participation of MCs as a factor of the tumor microenvironment in the PCa progression.