Russian Journal of BiotherapyRussian Journal of BiotherapyРоссийский биотерапевтический журнал1726-97841726-9792Publishing House ABV Press115510.17650/1726-9784-2019-18-2-40-50ORIGINAL REPORTSОРИГИНАЛЬНЫЕ СТАТЬИStudy of antitumor activity of synthetic peptide ryqlhpyr on the prostate cancer cellsИзучение противоопухолевой активности синтетического пептида RYQLHPYR на клетках рака предстательной железыhttps://orcid.org/0000-0002-9126-3070AkentievaN. P.АкентьеваН. П.
Russian Federation

Prospect Akademika Semenova, Chernogolovka 142432; 1 Prospect Akademika Semenova, Chernogolovka 142432.

42432 Черноголовка, пр-т акад. Семенова, 1; 142432 Черноголовка, пр-т акад. Семенова, 1.

na_aken@icp.ac.ruhttps://orcid.org/0000-0002-2273-3024ShushanovS. S.ШушановС. С.
Russian Federation

24 Kashyrskoe Sh., Moscow 115478.

115478 Москва, Каширское ш., 24.

Institute of Problems of Chemical Physics, Russian Academy of SciencesФГБУН «Институт проблем химической физики РАН»Scientific-educational center “Medical chemistry” of the Moscow State Regional University.Научно-образовательный центр «Медицинская химия» ГОУ ВО МО «Московский государственный областной университет».N.N. Blokhin National Medical Research Center of Oncology of the Ministry of Health of Russia.ФГБУ «Национальный медицинский исследовательский центр онкологии им. Н. Н. Блохина» Минздрава России.1506201918240501506201915062019https://bioterapevt.abvpress.ru/jour/article/view/1155

Introduction . The RHAMM (hyaluronan mediated mobility receptor) is overexpressed in many types of human cancer and increased synthesis of the RHAMM usually correlates with a poor prognostic factor. In this paper, we synthesized the peptide-RYQLHPYR modulating the activity of the RHAMM and examined the therapeutic potential of this RHAMM-targeting peptide as an antitumor agent.

Objective . Study the effect of the synthetic peptide RYQLHPYR on viability, apoptosis, necrosis, caspase-3 / 7 activity, and invasion of prostate cancer cells.

Materials and methods . The peptide RYQLHPYR was prepared by solid phase synthesis. Human prostate cancer cells (PC3 m-LN4), murine embryonic fibroblasts and murine embryonic fibroblasts (RHAMM- / -). To quantify the effect of the peptide on apoptosis and cell necrosis, ELISAPLUS was used. The activity of caspase-3 / 7 was determined by the colorimetric method. Evaluation of the anti-metastatic effect of the peptide in vitro was evaluated by invasion of cells by quantitative analysis of the area of degradation of fluorescent gelatin.

Results . It was found that the peptide RYQLHPYR inhibited the growth of tumor cells PC3 m-LN4 at a concentration of 10 μg / ml (2 × 10–7 M) after 24 h by ~80 %. It was shown that the peptide stimulated the level of apoptosis in cancer cells, approximately 10-fold. It was found that the peptide increased the necrotic death of tumor cells by 2.5 times. During the research it was revealed that the peptide increased the caspase-3 / 7 activity in tumor cells by 2 times. At the same time, it was shown that RHAMM-targeting peptide had no significant effect on apoptosis and necrosis of normal cells (fibroblasts) and fibroblasts (RHAMM- / -). It was found that the peptide inhibited invasion of tumor cells by ~99.86 % at a concentration of 10 μg / ml (2 × 10–7 M).

Conclusions . The obtained results indicate that the peptide RYQLHPYR has antitumor activity and, therefore, has a therapeutic potential for the treatment of prostate cancer. 

Введение . RHAMM (опосредованный гиалуронаном рецептор подвижности) избыточно экспрессируется во многих типах рака человека, а повышенный синтез RHAMM обычно коррелирует с плохим прогностическим фактором. В ходе исследования мы синтезировали пептид RYQLHPYR, модулирующий активность RHAMM, и рассмотрели терапевтический потенциал этого RHAMM-таргет-пептида как противоопухолевого реагента.

Цель исследования – изучение влияния синтетического пептида RYQLHPYR на жизнеспособность, апоптоз, некроз, активность каспаз-3 / 7 и инвазивность клеток рака предстательной железы.

Материалы и методы . Пептид RYQLHPYR был получен твердофазным синтезом. Изучались клетки рака предстательной железы человека (PC3 m-LN4 ), мышиные эмбриональные фибробласты (неинвазивные, нормальные клетки) и мышиные эмбриональные фибробласты (RHAMM- / -). Для количественной оценки влияния пептида на апоптоз и некроз клеток использовали метод ELISAPLUS. Определение активности каспаз-3 / 7 проводили колориметрическим методом. Антиметастатическое действие пептида in vitro оценивалось по инвазивности клеток методом количественного анализа площади деградации флуоресцентного желатина.

Результаты . Установлено, что пептид RYQLHPYR ингибировал рост опухолевых клеток PC3 m-LN4 при концентрации 10 мкг / мл (2 × 10–7 М) через 24 ч на ~80 %. Показано, что пептид стимулировал повышение уровня апоптоза в раковых клетках примерно в 10 раз и ускорял некротическую гибель опухолевых клеток в 2,5 раза. В ходе исследований выявлено, что пептид повышал в 2 раза активность каспаз-3 / 7 в опухолевых клетках. В то же время показано, что RHAMM-таргетпептид не оказывал значительного эффекта на апоптоз и некроз нормальных клеток и RHAMM- / -. Установлено, что пептид ингибировал инвазивность опухолевых клеток на ~99,86 % при концентрации 10 мкг / мл (2 × 10–7 М).

Выводы . Полученные результаты указывают, что пептид RYQLHPYR обладает противоопухолевой активностью и, следовательно, имеет терапевтический потенциал для лечения рака предстательной железы. 

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