Tumors of female reproductive systemTumors of female reproductive systemОпухоли женской репродуктивной системы1994-40981999-8627Publishing House ABV Press42310.17650/1994-4098-2015-1-61-68MAMMOLOGY. DIAGNOSISМАММОЛОГИЯ. ДИАГНОСТИКАANALYSIS OF APOPTOSIS FACTORS IN PATIENTS WITH PREINVASIVE AND INVASIVE CERVICAL CANCERИССЛЕДОВАНИЕ ПОКАЗАТЕЛЕЙ АПОПТОЗА У БОЛЬНЫХ С ПРЕИНВАЗИВНЫМ И ИНВАЗИВНЫМ РАКОМ ШЕЙКИ МАТКИKovchurP. I.КовчурП. И.
Russian Federation
pkovchur@mail.ruBachidzeE. V.БахидзеЕ. В.
Russian Federation
KurmyshkinaO. V.КурмышкинаО. В.
Russian Federation
VolkovaT. O.ВолковаТ. О.
Russian Federation
Petrozavodsk State UniversityФГБОУ ВПО «Петрозаводский государственный университет»33 Lenina prospekt, Petrozavodsk, 185910, RussiaРоссия, 185910, Петрозаводск, проспект Ленина, 33N.N. Petrov Oncology Research Institute, Ministry of Health of RussiaФГБУ «Научно-исследовательский институт онкологии им. Н.Н. Петрова» Минздрава России68 Leningradskaya St., Pesochny, Saint Petersburg, 197758, RussiaРоссия, 197758, Санкт-Петербург, пос. Песочный, ул. Ленинградская, 682904201511161683004201530042015https://ojrs.abvpress.ru/ojrs/article/view/423

This article assesses the disturbance degree of apoptotic program in patients with preinvasive and invasive cervical cancer by investigating the expression level of genes of caspases-3, -6, -8 и -9 in mononuclear cells of periphery blood and in tumor tissue on the two regulating levels – on mRNA level (transcriptional) and proteolytic activity (post transcriptional). 75 patients with stage III of cervical intraepithelial neoplasias (CIN) III  (middle age of 32,9 ± 7,4),45 patients with stage IA (31,3 ± 6,0), 21 – with stage II (43,6 ± 13,2), 15 – with stage III–IV (46,9 ± 11,1) have been examined. The control group has been formed from 30 almost healthy donors without any cervical pathology and papilloma human virus (control 1) and 30 patients with a preinvasive and microinvasive cervical cancer (control 2). It has been found that in proportion of progress of cervix cancer, the membranous expression of CD95 increases in MPB – fraction (peripheral blood monocytes) when the a CIN and initial stages of cervix cancer, more than two times. Herewith number CD95+-lymphocytes is positively correlated with stage of cervical cancer (r = 0,91; R2  = 0,82; p << 0,01). It has been found out that the activity gain of caspase-8 (r = 0,92; R2  = 0,86; p << 0,01), caspase-6 (r = 0,77; R2 = 0,59; p << 0,01) and reduction activity of caspase-9 (r = –0,60;  R2 = 0,36; p < 0,01) in mononuclear cells of peripheral blood pointed out on the sensitivity increase to Fas-induced apoptosis. Opposite, in tumor tissue, beginning from CIN stage III, apoptosis-resistant phenotype is formed, it were defined by the expression of caspase-3 (r = –0,72; R2 = 0,52, p < 0,01), caspase-6 (r = –0,59; R2 = 0, 38; p < 0,01) и caspase-9 (r = –0,67; R2 = 0,45; p < 0,01) by mRNA level and proteolytic activity. It has been shown, that the cervical cancer development is accompanied by multilateral disturbances of apoptotic processes, which are realized in decreased function of caspases and multilateral resistance of tumor cells in against signals of apoptosis. The obtained results give an opportunity to examine the genes of caspase as a probable prospective biomarkers in complex diagnostic of a CIN and early diagnosis of a cervical cancer (in combination with morphological criteria and other molecular markers of viral origin)

В статье дана оценка степени нарушения апоптотической программы у больных преинвазивным и инвазивным раком шейки матки

(РШМ) с помощью определения экспрессии генов каспаз-3, -6, -8 и -9 в мононуклеарах периферической крови (МНПК) и опухолевой ткани на двух уровнях регуляции – мРНК (транскрипционный уровень) и протеолитической активности (посттрансляционный уровень). Обследованы 75 больных с цервикальными интраэпителиальными неоплазиями (ЦИН) III степени тяжести (средний возраст 32,9 ± 7,4 года), 45 – с IА стадией (31,3 ± 6,0 года), 21 – со II стадией (43,6 ± 13,2 года), 15 – с III–IV стадиями (46,9 ±

11,1 года) РШМ. Были выделены 2 контрольные группы: в 1-ю вошли 30 практически здоровых доноров без патологии шейки матки и вируса папилломы человека (ВПЧ), во 2-ю – 30 больных преинвазивным и микроинвазивным РШМ. Установлено, что по мере прогрессии РШМ происходит повышение мембранной экспрессии CD95 в МНПК-фракции при ЦИН III степени и ранних стадиях РШМ более чем в 2 раза и наблюдается положительная корреляция количества CD95+-лимфоцитов со стадией РШМ (r = 0,91; R2 = 0,82; p << 0,01). Обнаружено усиление активности каспазы-8 (r = 0,92; R2 = 0,86; p << 0,01), каспазы-6 (r = 0,77; R2 = 0,59; p << 0,01) и подавление активности каспазы-9 (r = –0,60; R2 = 0,36; p < 0,01) в МНПК, указывающие на повышение чувствительности к Fas- индуцированному апоптозу. Напротив, в ткани опухоли, начиная с ЦИН III степени, формируется апоптоз-резистентный фенотип, что определено с помощью экспрессии каспазы-3 (r = –0,72; R2 = 0,52; p < 0,01), каспазы-6 (r = –0,59; R2 = 0,38; p < 0,01) и каспазы-9 (r = –0,67; R2 = 0,45; p < 0,01) по уровню мРНК и протеолитической активности. Показано, что развитие РШМ сопровождается многосторонними нарушениями апоптотических процессов, которые реализуются в подавлении функционирования каспаз и множественной резистентности опухолевых клеток к апоптотическим стимулам. Полученные результаты дают возможность рассматривать гены каспазы как возможные перспективные биомаркеры в комплексной диагностике ЦИН и ранних форм РШМ (в сочетании с морфологическими критериями и другими молекулярными маркерами вирусного происхождения).

 

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