Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells

Aim - the aim of the study was in vitro analysis of biological activity of recombinant human beta-defensin-4 (rec-hBD-4). Methods: hBD-4 cDNA was cloned into pGEX-2T vector, and recombinant plasmid was transformed into E. coli BL21(DE3) cells. To purify soluble fusion GST-hBD-4 protein, affinity chromatography was applied. Rec-hBD-4 was cleaved from the fusion protein with thrombin, and purified by reverse phase chromatography on Sep-Pack C18. Effects of rec-hBD-4 on proliferation, viability, cell cycle distribution, substrate-independent growth, and mobility of cultured human cancer cells of A431, A549, and TPC-1 lines were analyzed by direct cell counting technique, MTT assay, flow cytofluorometry, colony forming assay in semi-soft medium, and wound healing assay. Rech-BD-4 was expressed in bacterial cells as GST-hBD-4 fusion protein, and purified by routine 3-step procedure (affine chromatography on glutathione-agarose, cleavage of fusion protein by thrombin, and reverse phase chromatography). Analysis of in vitro activity of rec-hBD-4 toward three human cancer cell lines has demonstrated that the defensin is capable to affect cell behaviour in concentration-dependent manner. In 1 - 100 nM concentrations rec-hBD-4 significantly stimulates cancer cell proliferation and viability, and promotes cell cycle progression through G2/M checkpoint, greatly enhances colony-forming activity and mobility of the cells. Treatment of the cells with 500 nM of rec-hBD-4 resulted in opposite effects: significant suppression of cell proliferation and viability, blockage of cell cycle in G1/S checkpoint, significant inhibition of cell migration and colony forming activity

The influence of GSTP1 A313G polymorphism on susceptibility, chemotherapy-related toxicity and prognosis of Hodgkin’s lymphoma in Ukrainian patients

The aim of this research was to study the influence of GSTP1 A313G polymorphism on susceptibility, chemotherapy-related toxicity and prognosis of Hodgkin’s lymphoma in Ukrainian patients. Methods. The polymorphic variants of GSTP1 gene were analyzed using Allelic Discrimination Real-Time PCR. Results. The GSTP1 polymorphism is not directly involved in the development of Hodgkin’s lymphoma and chemotherapy-related toxicity, but homozygous wild genotype of this gene is associated with a worse clinical response to the therapy and a higher risk of relapse. Conclusions. The investigation of GSTP1 polymorphism is very promising, since it might provide a possible application of this genetic marker as an independent prognostic factor of Hodgkin’s lymphoma.Мета. Дослідити поліморфізм гена GSTP1 A313G і визначити його вплив на виникнення лімфоми Ходжкіна, токсичність хіміотерапії, а також на перебіг захворювання. Методи. Поліморфні варіанти гена GSTP1 аналізували методом алель-специфічної ПЛР з детекцією результатів у режимі реального часу. Результати. Встановлено, що гомозиготний тип успадкування алеля дикого типу – генотип A/A – пов’язаний з несприятливим прогнозом перебігу лімфоми Ходжкіна: гіршою відповіддю на терапію і вірогідністю появи рецидивів. Висновки. Перспективним напрямком пошуку чинників впливу на перебіг хвороби є вивчення поліморфізму гена GSTP1, який у майбутньому може бути використаний як «фактор ризику» при формуванні прогностичних груп хворих та виборі тактики лікування.Цель. Исследовать полиморфизм гена GSTP1 A313G и определить его влияние на возникновение лимфомы Ходжкина, токсичность химиотерапии, а также на течение заболевания. Методы. Полиморфные варианты гена GSTP1 анализировали методом аллель-специфической ПЦР с детекцией результатов в режиме реального времени. Результаты. Установлено, что гомозиготный тип наследования аллеля дикого типа – генотип A/A – ассоциирован с неблагоприятным прогнозом течения лимфомы Ходжкина: худшим ответом на лечение и вероятностью появления рецидивов. Выводы. Перспективным направлением поиска причин, обусловливающих течение болезни, является изучение полиморфизма гена GSTP1, который в будущем, возможно, будет использован как «фактор риcка» при формировании прогностических групп больных и выборе тактики лечения

Results from phase III clinical trials with radachlorine for photodynamic therapy of pre-cancer and early cancer of cervix

The results of clinical study for efficacy of photodynamic therapy (PDT) with radachlorine in patients with pre-cancer and cancer of cervix are represented. The study enrolled 30 patients including 4 patients with cervical erosion, 5 patients with cervical intraepithelial neoplasia II, 13 patients with cervical intraepithelial neoplasia III, 4 patients with carcinoma in situ and 4 patients with cervical cancer stage Ia. Radachlorine was administrated as single 30 minute intravenous injection at dose of 1,0 mg/kg of body weight 3 h before irradiation (wavelength of 662 nm, light dose of 300-350 J/cm2). The results of treatment in 26 (86,7%) patients was assessed as complete tumor regression and in 4 (13,3%) patients - as partial regression. In cervical erosion, intraepithelial neoplasia II and carcinoma in situ groups total regression was in all cases. In the cervical intraepithelial neoplasia III group total regression after first course of PDT was achieved in 77% of patients, in cervical cancer stage Ia group - in 75% of patients. From 3 to 6 months after first course of treatment all patients with partial tumor regression underwent the second course of PDT with complete regression. There were no side-effects due to radachorine or PDT in the course of treatment and during follow-up. Thus, PDT with Russian photosensitizer radachlorine showed high efficiency for treatment of pre-cancer and cancer of cervix

Capabilities of intraoperative photodynamic therapy for treatment of locally advanced breast cancer

The original method of intraoperative photodynamic therapy for multimodality treatment of primary and recurrent breast cancer for devitalization of malifnant cells at wound surface and for prevention of further cancer dissemination was developed in P.A. Herzen Moscow Cancer Research Institute. The developed method was approved in 79 patients with locally advanced breast cancer stage IIB and IIIA,B,C with poor prognostic factors. For photodynamic therapy the photosensitizer photosens (30 min intravenous infusion at dose of 0.3 mg/kg of body weight 2 h before surgery) was used in 56 patients; alasens (solution in 100 ml of still drinking water, orally at dose of 30 mg/kg of body weight 2 h before general anesthesia) - in 23 patients. The surgical field was irradiated on a single occasion: the dose of laser irradiation on the bed of removed primary or recurrent tumor was 20-30 J/cm2, in the removed regional lymph nodes area - 50 J/cm2. Long-term results of the treatment were assessed in 34 patients: there were no disease progression in 50% of patients, 14.7% of patients had locoregional recurrence, distant metastases were in 35.3% of patients. The level of photosensitizer accumulation in tissue was additionally analyzed in 26 patients by fluorescence intensity in tumor and in normal breast tissue. After injection of alasens the increase in level of alasensinduced protoporphyrin IX fluorescence was recorded in tumor (the average diagnostic parameter was 6.5 a.u.) and in intact breast tissue (an average of 0.47 a.u.), tumor/normal tissue fluorescence contrast varied from 3 to 33. The level of protoporphyrin IX accumulation was noticed to be lower in tumors with pathomorphological response after neoadjuvant therapy. For photosens value of the average diagnostic parameter in normal breast tissue was 5.6 a.u., in tumor - 34.3 a.u., tumor/normal tissue fluorescence contrast - from 2 to 9

PHOTODYNAMIC THERAPY OF A PATIENT WITH BASAL CELL SKIN CANCER OF THE EAR STAGE T3N0M0(CLINICAL CASE) [ФОТОДИНАМИЧЕСКАЯ ТЕРАПИЯ БОЛЬНОГО БАЗАЛЬНОКЛЕТОЧНЫМ РАКОМ КОЖИ УШНОЙ РАКОВИНЫ СТАДИИ T3N0M0 (КЛИНИЧЕСКОЕ НАБЛЮД ЕНИЕ)]

The article presents a clinical observation. The patient, 72 years old, applied to the MNII them. P.A. Herzen with complaints about the presence of an ulcerated tumor of the left ear. After further examination, a diagnosis was made - basal cell carcinoma of the ear skin with spread to the skin of the parotid region T3N0M0. On July 9, 2021, the patient underwent a course of photodynamic therapy (PDT) using a photosensitizer based on chlorin e6 and a diode laser with a wavelength of 662 nm. After one course of PDT, complete regression of the tumor was recorded. © 2021 Russian Photodynamic Association. All rights reserved

Photodynamic therapy for facial skin cancer developed in the zone of previous radiotherapy (clinical case) [ФОТОдИНАМИчЕСКАя ТЕРАПИя ПРИ РАКЕ КОЖИ ЛИЦА, РАЗВИВШЕГОСя В ЗОНЕ ПРЕдШЕСТВУюЩЕЙ ЛУчЕВОЙ ТЕРАПИИ (КЛИНИчЕСКОЕ НАбЛюдЕНИЕ)]

The results of a 13-year clinical observation of a patient after treatment for basal cell carcinoma of the skin of the right cheek Ist cT1N0M0 are presented. The history of the course of the disease is associated with the fact that the patient underwent radiation therapy in early childhood for hemangioma of the lower eyelid of the right eye and right cheek. In 2008, against the background of post-radiation changes in the area of the right cheek, basal cell carcinoma was diagnosed at the Moscow Oncological Research Institute. P.A. Herzen. At the Center for Laser and Photodynamic Diagnostics and Tumor Therapy, the patient underwent organ-preserving PDT treatment. A course of photodynamic therapy (PDT) with 5-aminolevulic acid was carried out. Subsequently, the patient was followed up until 2021 without relapse in the PDT area. In 2016, the patient was diagnosed with a relapse of the disease in the form of a new focus of basal cell carcinoma of the upper eyelid skin on the right Iast cT1N0M0. The patient underwent a course of PDT with a chlorin e6-based photosensitizer. Complete regression of the tumor was achieved, the period of relapse-free follow-up was 5 years. © 2021 Russian Photodynamic Association. All rights reserved

Results from phase III clinical trials with radachlorine for photodynamic therapy of pre-cancer and early cancer of cervix

The results of clinical study for efficacy of photodynamic therapy (PDT) with radachlorine in patients with pre-cancer and cancer of cervix are represented. The study enrolled 30 patients including 4 patients with cervical erosion, 5 patients with cervical intraepithelial neoplasia II, 13 patients with cervical intraepithelial neoplasia III, 4 patients with carcinoma in situ and 4 patients with cervical cancer stage Ia. Radachlorine was administrated as single 30 minute intravenous injection at dose of 1,0 mg/kg of body weight 3 h before irradiation (wavelength of 662 nm, light dose of 300-350 J/cm2). The results of treatment in 26 (86,7%) patients was assessed as complete tumor regression and in 4 (13,3%) patients - as partial regression. In cervical erosion, intraepithelial neoplasia II and carcinoma in situ groups total regression was in all cases. In the cervical intraepithelial neoplasia III group total regression after first course of PDT was achieved in 77% of patients, in cervical cancer stage Ia group - in 75% of patients. From 3 to 6 months after first course of treatment all patients with partial tumor regression underwent the second course of PDT with complete regression. There were no side-effects due to radachorine or PDT in the course of treatment and during follow-up. Thus, PDT with Russian photosensitizer radachlorine showed high efficiency for treatment of pre-cancer and cancer of cervix

Capabilities of intraoperative photodynamic therapy for treatment of locally advanced breast cancer

The original method of intraoperative photodynamic therapy for multimodality treatment of primary and recurrent breast cancer for devitalization of malifnant cells at wound surface and for prevention of further cancer dissemination was developed in P.A. Herzen Moscow Cancer Research Institute. The developed method was approved in 79 patients with locally advanced breast cancer stage IIB and IIIA,B,C with poor prognostic factors. For photodynamic therapy the photosensitizer photosens (30 min intravenous infusion at dose of 0.3 mg/kg of body weight 2 h before surgery) was used in 56 patients; alasens (solution in 100 ml of still drinking water, orally at dose of 30 mg/kg of body weight 2 h before general anesthesia) - in 23 patients. The surgical field was irradiated on a single occasion: the dose of laser irradiation on the bed of removed primary or recurrent tumor was 20-30 J/cm2, in the removed regional lymph nodes area - 50 J/cm2. Long-term results of the treatment were assessed in 34 patients: there were no disease progression in 50% of patients, 14.7% of patients had locoregional recurrence, distant metastases were in 35.3% of patients. The level of photosensitizer accumulation in tissue was additionally analyzed in 26 patients by fluorescence intensity in tumor and in normal breast tissue. After injection of alasens the increase in level of alasensinduced protoporphyrin IX fluorescence was recorded in tumor (the average diagnostic parameter was 6.5 a.u.) and in intact breast tissue (an average of 0.47 a.u.), tumor/normal tissue fluorescence contrast varied from 3 to 33. The level of protoporphyrin IX accumulation was noticed to be lower in tumors with pathomorphological response after neoadjuvant therapy. For photosens value of the average diagnostic parameter in normal breast tissue was 5.6 a.u., in tumor - 34.3 a.u., tumor/normal tissue fluorescence contrast - from 2 to 9