Experimental oncology最新文献

Background: The level of heat shock protein 60 (Hsp60) is elevated in tumor cells compared with normal prostate epithelium. Hsp60 is involved in tumor growth, invasion, and metastasis and is considered as a biomarker for cancer diagnosis and prognosis.

Aim: To study the level of antibodies against prokaryotic homolog of human Hsp60 (GroEL) in prostate cancer (PCa) patients as an additional risk marker for the prediction of biochemical recurrence after radical prostatectomy (RP).

Patients and methods: A total of 55 patients with localized and locally advanced PCa, who had undergone RP between July 2013 and May 2014 were enrolled. Level of antibodies to GroEL and human Hsp60 was determined by enzyme-linked immunosorbent assay before surgery. Serum samples of blood donors with low reactivity to GroEL and human Hsp60 were used as controls. The relationship between IgG antibodies against bacterial Hsp60 and human Hsp60 and clinicopathological features were analyzed. The biochemical recurrence (BCR) free survival rate was estimated by the Kaplan - Meier method. The univariate and multivariate Cox regression models were used to evaluate the risk factors of BCR-free survival rate.

Results: There were significant differences in anti-GroEL IgG levels between control and PCa patients while no significant differences in anti-human Hsp60 IgG levels between control and PCa patients were detected. During the follow-up period, 40/55 (72.7%) patients developed BCR. The time from surgery to BCR was from 18 to 72 months. Elevated IgG antibodies against bacterial Hsp60 in patients who had undergone RP were associated with early occurrence of biochemical relapse and lower 5-year BCR-free survival rate respectively (p < 0.001). The multivariate analysis indicated that IgG to GroEL (hazard ratio = 2.465; 95% confidence interval: 1.311-4.634, p < 0.05) could be independent prognostic factor in the patients who had developed BCR.

Conclusion: Elevated levels of IgG antibodies against GroEL before surgery can predict early occurrence of BCR after RP and can serve as an additional independent risk biomarker of a BCR after RP.

Background: According to modern literature, osteopontin (OPN) and osteonectin (ON) are involved not only in the formation of the aggressive phenotype of malignantly transformed cells, but also in the realization of cytotoxic effects of some antitumor drugs.

Aim: To study the changes of the expression of OPN and ON and their mRNAs (SPP1 and SPARC) upon exposure to doxorubicin (Dox) in breast cancer (BCa) and prostate cancer (PCa) cell lines with different sensitivity to Dox.

Materials and methods: Cell lines of BCa (MCF-7 and MDA-MB-231) and PCa (LNCaP and DU-145) were cultured in the presence of Dox at IC30 concentrations for 24 h. OPN and ON levels were assessed by immunocytochemical (ICH) and Western blot analysis. SPP1 and SPARC mRNA levels were assessed by quantitative PCR.

Results: Dox treatment resulted in the significant decrease in the expression of both OPN and ON in MCF-7 and LNCaP cells. Similarly, Dox treatment downregulated both SPP1 and SPARC in MDA-MB-231 and DU-145 cells. Dox did not affect ON expression in MDA-MB-231 and DU-145 cells although the significant decrease in the level of SPARC mRNA has been evident. In contrast, no significant differences in SPP1 and SPARC mRNA levels were detected in LNCaP cells.

Conclusion: The changes in the expression of OPN and ON proteins and their corresponding genes in BCa and PCa cells may be related to the intrinsic mechanisms of Dox effects in cells differing by malignant phenotype and Dox sensitivity.

Background: Hypoxia has been noted as a key factor for induction and maintenance of cancer stemness thereby leading to therapy resistance. Three-dimensional (3D) spheroid models demonstrate a heterogeneity of hypoxic regions replicating the in vivo situation within tumors. Utilizing an established 3D spheroid model, we investigated whether extrinsic hypoxia reinforced chemoresistance in malignant pleural mesothelioma (MPM) spheroids.

Materials and methods: Tumor spheres were generated from Meso-1 (a typical human MPM cell line) cells having high spheroid-forming ability. To induce hypoxia condition, we utilized a hypoxia chamber with regulation of O2 and CO2 levels. Cell viability was estimated by a WST-8 assay. Real-time polymerase chain reaction and Western blot were performed to evaluate the expression at mRNA and protein levels.

Results: Compared with cells cultured in the two-dimensional monolayer model, tumor sphere cells showed elevated mRNA levels of cancer stemness markers (CD26, CD44 and ABCG2) and protein levels of the stemness and hypoxia adaptation markers (ABCG2, ALDH1A1 and HIFs). Correlating with this, 3D spheroid cells were more resistant to permetrexed and topotecan than the two-dimensional cells, indicative of their potential for hypoxic adaptation. Furthermore, significantly stronger resistance to both chemotherapeutic agents was observed in spheroid cells upon hypoxic challenge compared to spheroid cells under normoxia.

Conclusion: From the present data, it is concluded that hypoxia adaptation of MPM cells from tumor spheres could enhance their chemoresistance.

Oncolytic virotherapy is an emerging biotherapeutic platform for selectively infecting cancer cells and triggering apoptosis in a number of malignant cells due to robust viral replication. Studies related to the oncolytic activity of human orthopneumovirus (hOPV) are conflicting.

Aim: This study was designed to elucidate the possible role of hOPV in the modulation of cell growth and apoptosis in cancer cell lines including human epidermoid carcinoma (HEp-2), lung epithelial cell line (A549), and breast cancer cell line (MCF-7).

Materials and methods: The oncolytic activity of hOPV on cancer cells was studied in vitro. The virus titers were determined by tissue culture infectious dose (TCID50/mL) in A549 cell. The cytotoxic effect of the virus on HEp-2, A549, and MCF-7 was determined using MTT and trypan blue dye exclusion test assays. hOPV in the infected cells was detected using real-time reverse transcription polymerase chain reaction (rRT-PCR) and indirect immunofluorescence (IIF) assays. The relative expression of apoptosis-related genes (CASP-3, -8, -9, Bax, Bcl-2, Bcl-XL, TP53, P21) during virus infection was estimated using rRT-PCR assay in comparison with the house-keeping gene (GAPDH).

Results: hOPV infection inhibited the growth of HEp-2, A549, and MCF-7 cells in a dose-and time-dependent manner. At a multiplicity of infection (MOI) of 5, hOPV reduced the viability of A549 cells to about 16%, HEp-2 to 22%, and MCF-7 to 28% (p = 0.001), while no significant inhibitory effect was observed when cells were infected at MOI of 1 and 2. hOPV mRNA and antigens were detected in infected HEp-2, A549, and MCF-7 cells by RT-PCR and IIF. Upon hOPV infection, expression of CASP-3, -8, -9, as well as Bax, TP53, and p21 mRNA increased while expression of Bcl-2, Bcl-xL anti-apoptotic genes decreased. In hOPV-infected A549 cells, the fold increase of CASP-8 and CASP-9, Bax, TP53, and P21 expression exceeded significantly compared to that in HEp-2 or MCF-7 cells.

Conclusions: Our results provide evidence that hOPV could be a potential candidate for oncolytic virotherapy.

Background: Metastases in eye structures are rare (1-5% cases at systemic spread of different malignancies, mainly breast and lung cancers). The prognosis is poor. The overall survival usually does not exceed 12 or even 6 months. If metastases are found in the choroid membrane, the probability that the patient has multiple metastatic lesions of other organs increases significantly. Lung neuroendocrine neoplasms are rare (1-2% of all malignancies in adults), but mainly aggressive tumors. They are cha-racterized by "blurred", nonspecific clinical symptoms, the correct diagnosis is delayed seriously, and distant metastases are seen in more than 40% of patients (usually in chest structures, liver, bones, brain, and adrenal glands; metastasis to vascular membrane of the eye ranks the 6th place).

Case report: Own clinical observation of a male patient with rare metastasis of lung neuroendocrine carcinoma to the choroid of the left eye is presented. The disease is manifested by an ocular metastasis, which was initially considered an embryonic tumor. Other metastatic lesions (hilar lymph nodes, liver, soft tissues) were detected on computed tomography a little bit later. The diagnostic algorithm using routine histological examination and immunohistochemistry, including detection of neuroendocrine markers (chromogranin A, synaptophysin), cytokeratin 7 and Ki-67 expression in primary and metastatic tumors is described.

Background: In spite of significant advances in diagnosis of prostate cancer (PCa), the detection and differential diagnosis of metastatic lymph node involvement remains an important clinical dilemma in a large number of cases. Contrast-enhanced abdominal computed tomography and magnetic resonance imaging (MRI), in part when using T1-weighed images (T1-WI and T2-WI), allow evaluating indirectly the presence of invasion in regional lymph nodes by assessing their diameter and morphology. Nonetheless, these techniques do not appear to be sufficiently sensitive for direct identification of lymph nodes with metastatic lesions.

Aim: To study the significance of the apparent diffusion coefficient (ADC) of diffusion-weighted MRI in detection of metastatic lymph node involvement in PCa patients.

Materials and methods: The study involved 35 patients with histologically verified PCa. Based on multiparametric prostatic MRI findings and pathomorphological reports, we have performed ADC measurements for pelvic lymph nodes either with (n = 15, mean size 1.78 ± 0.59 cm) or without metastases (n = 20, mean size: 0.94 ± 0.06 cm) in PCa patients who underwent radical prostatectomy with lymph node dissection.

Results: No significant diffe-rences were observed when comparing mean sizes of N+ and N- pelvic lymph nodes. At the same time, when comparing mean ADC values for N+ and N- pelvic lymph nodes, we observed a statistically significant difference: 0.74 ± 0.09 · 10-3 mm2/s in metastatic lymph node vs 1.05 ± 0.23 · 10-3 mm2/s in lymph nodes without metastatic involvement (p < 0.001).

Conclusion: The use of ADC for diffusion-weighted MRI may provide valuable information for detection of metastatic lymph node involvement in patients with PCa.

Background: Chemokine (C-C motif) receptor 7 (CCR7) is a chemokine receptor involved in the carcinogenesis of several types of tumors due to its promoting action in epithelial-mesenchymal transition events, invasion, angiogenesis and metastasis. However, its role in prostate cancer (PCa) remains unclear.

Aim: To evaluate CCR7 expression by immunohistochemistry in prostate tumors from young patients and to determine the possible relationship with the clinicopathological characteristics.

Materials and methods: We analyzed retrospectively paraffin-embedded tissue sections from 23 young PCa (≤ 55 years old) patients and evaluated the transcriptomic expression in the TCGA database.

Results: Expression of CCR7 was observed in 15 cases (65%). The tissue samples from younger patients (≤ 50 years) were mostly positive in 72.7% (8/11) of cases. High grade GS (≥ 3) tumors were CCR7-positive in 71% cases. The malignant cells present in lymph nodes were CCR7 positive in 100% cases. The bioinformatic analysis showed a high CCR7 expression associated with the presence of metastasis (FC = 2.6, p = 0.03) in the Cancer Genome Atlas (TCGA) PCa cohort (PRAD).

Conclusion: We showed that CCR7 expression in tumors from young patients is associated with the early onset of the disease and could also be related to lymph node metastasis.

Histone deacetylases, especially zinc-dependent deacetylases HDACs, are among attractive drug targets for treating cancer in recent years.

Aim: To explore the expression level of HDACs in several human cancer cell lines and examine the possible association between their expression and the sensitivity/resistance to the selective- or pan-HDAC inhibitors.

Materials and methods: The RNA expression of 11 HDACs isoforms was assayed in HeLa, HepG2, AV3, HEK293, A549, and K562 cells by semiquantitative reverse transcription-polymerase chain reaction. The sensitivity/resistance of these cell lines to the pan- or selective- HDAC inhibitors was estimated by MTS assay.

Results: The relative transcription of HDACs genes demonstrated that members of Class I HDAC (HDAC1, 2 and 3) and members of Class II HDAC (HDAC4, 5, 6 and 7) had slight to significant levels of expression in cell lines under study with no dominant HDAC-subtype gene transcription. pan-HDAC inhibitor demonstrated superior antitumor activity compared to HDAC isoform-selective inhibitor.

Conclusion: The absence of the dominant HDAC-subtype gene transcription in different human cancer cell lines explains the inferior efficacy of HDAC isoform-selective inhibitors as compared to pan-HDAC inhibitors.