Research progress on treatment strategies for related diseases based on Parthanatos / 中国生物制品学杂志

@#Parthanatos is a form of programmed cell death,which is also known as poly(ADP-ribose)polymerase 1(PARP1)-mediated apoptosis-inducing factor(AIF)and macrophage migration inhibitory factor(MIF)-dependent cell death according to its molecular mechanism. Parthanatos is the main cause of a variety of neurodegenerative diseases,such as Parkinson's disease(PD),Alzheimer's disease(AD),motor neuron disease,and is also involved in the pathogenesis of some tumors,such as lung cancer and breast cancer. Therefore,a thorough understanding of the molecular mechanism of Parthanatos is crucial for the therapeutic strategies of related diseases. In recent years,studies have found that effective regulation of the occurrence of Parthanatos by regulating the key proteins PARP1,AIF and MIF is expected to become a therapeutic target for many diseases. Based on the specific molecular mechanism of Parthanatos,this paper reviewed the research progress of therapeutic strategies for related diseases from the aspects of inhibiting and promoting Parthanatos.

Research progress on the involvement of poly (ADP-ribose) polymerase-1-dependent cell death in ischemia-reperfusion injury / 中国医师杂志

Ischemia reperfusion injury (IRI) of organs is a major challenge for clinicians, but the mechanism is still not elucidated, and the clinical treatment effect is still unsatisfactory. PARP-1-dependent cell death (parthanatos) is a non-apototic programmed cell death pathway involved in the development of the occurrence of IRI of organs. At the same time, parthanatos is also a multi-step pathway. There are many molecules in the parthanatos cascade that can be exploited to create therapeutic interventions for IRI, including PARP1, apoptosis inducing factor (AIF), and macrophage migration inhibitory factor (MIF). These critical molecules are involved in DNA damage, energy depletion and homeostasis imbalance. Therefore, these molecular signals in the parthanatos cascade represent promising therapeutic targets for the treatment of IRI. In the following, we will elaborate on the mechanisms and molecular characteristics of the parthanatos pathway and the relation between parthanatos pathway and IRI of vital organs. It aims to explore the posibility of IRI mechanism research and clinical treatment and to provide new ideas for clinicians and researchers.

Effect of acupuncture on neurological function, cerebral infarction volume, thrombolysis time window and cerebral cell apoptosis signaling pathway in cerebral infarction rats / 针刺研究

OBJECTIVE: To observe the effect of acupuncture (Acupunct) on cerebral infarction volume and expression of poly ADP ribose polymerase 1 (PARP1), apoptosis-inducing factor (AIF) and endonuclease G (Endo-G) in the cerebral cortex tissue at different time-points after cerebral ischemia (CI) in acute cerebral infarction rats, so as to explore its underlying mechanisms in prolonging time window of thrombolysis. METHODS: Forty-eight SD rats were randomly divided into sham operation, model, intravenous thrombolysis (IVT)-4.5 h, IVT-6 h, IVT-9 h, Acupunct＋IVT-4.5 h, Acupunct ＋IVT-6 h, Acupunct＋IVT-9 h groups (n＝6 in each group). The CI model was established by using modified autologous thromboembolism via the right common carotid artery. Two hours after modeling, rats of the Acupunct groups received Acupunct stimulation of "Shuigou" (GV26) and bilateral "Neiguan" (PC6) for 30 min. Thrombolysis was conducted by injection of recombinant human tissue-type plasminogen activator (rt-PA, 10 mg/kg) via caudal vein. The neurological deficit was assessed with reference to Bederson's methods. 2，3，5-triphenyltetrazolium chloride (TTC) staining was used to assess the cerebral infarction volume, and the expression of cerebral PARP1, AIF and Endo-G proteins detected by Western blot. RESULTS: Compared with the sham operation group, the neurological score and percentage of cerebral infarction volume, expression levels of PARP1, AIF and Endo-G proteins were significantly increased in the model group (P0.05). The levels of neurological score, percentage of cerebral infarction volume, and AIF expression were significantly lower in both the Acupunct＋IVT 4.5 h and Acupunct＋IVT-6 h groups than in the simple IVT-4.5 h and simple IVT-6 h groups, respectively (P<0.05), and the expression levels of PARP1 and Endo-G proteins were obviously lower in the Acupunct＋IVT-4.5 h group than in the IVT-4.5 h group (P<0.05). Endo-G proteins were obviously lower in the Acupunct＋IVT-9 h group than in the IVT-9 h group (P<0.05). CONCLUSION: Acupuncture may improve neurological function, reduce cerebral infarction volume and prolong the time window of thrombolysis in CI rats, which may be associated with its effect in suppressing AIF/PARP1/ Endo-G signaling.

Research progress of PARP-1 inhibitors in combination with other drugs to overcome drug resistance / 中国药科大学学报

@#Poly(ADP-ribose)polymerase-1(PARP-1)plays a vital role in the regulation of DNA repair and apoptosis. Breakthrough has been made in the treatment of cancer with PARP-1 inhibitors, but the emergence of drug resistance has limited its further application in clinic. This paper reviews advances in the research on PARP-1 inhibitors combined with other drugs to overcome drug resistance, highlights and evaluates the existing drug combination strategies and their therapeutic effects in clinical practice. It is proposed that the development of dual-target or multi-target drugs will become a promising approach to overcome the resistance of PARP-1 inhibitors and broaden their indications.

Carcinomas de ovário associados à endometriose: expressão de marcadores proteicos de terapia alvo e prognóstico / Endometriosis-associated ovarian carcinomas: expression of protein biomarkers associated to target therapies and prognosis

Resumo: Introdução: endometriose é uma doença benigna, capaz de progredir extensamente e gerar clones atípicos. Considerada precursora dos carcinomas de células claras (CCOC) e endometrióide (EOC) de ovário, atualmente chamados carcinomas de ovário associados à endometriose (EAOC). Objetivos: comparar o perfil epidemiológico, a associação com endometriose e a expressão de marcadores imuno-histoquímicos para ARID1A, VEGF, PD-L1 e PARP-1 em mulheres com CCOC e EOC, e sua correlação com a sobrevida livre de progressão (SLP) e sobrevida global (SG). Métodos: estudo de coorte reconstituída, com 50 casos incluídos de CCOC e EOC tratados no CAISM-UNICAMP entre 1995 até 2016, acompanhados até 02/2017. Microarranjos de tecido com amostras de CCOC, EOC e endometriose foram corados com anticorpos monoclonais contra ARID1A, e para os biomarcadores proteicos VEGF, PD-L1, PARP-1 através de imuno-histoquímica. A expressão de ARID1A foi classificada (0 a 100) conforme a porcentagem de células não coradas. A expressão de VEGF, PD-L1 e PARP-1 foi classificada (0 a 300) conforme a multiplicação da porcentagem de células coradas por um fator da intensidade de expressão (ausente=0; fraco=1; moderado=2; forte=3). Idade ao diagnóstico; menopausa; índice de massa corpórea (IMC); CA-125; diagnóstico de endometriose; datas do diagnóstico, da progressão, do óbito e da última consulta foram recuperados dos prontuários. Comparação entre grupos foi realizada através de testes T e de ?2. A SLP (diferença de tempo entre o diagnóstico e a data de progressão) e a SG (diferença de tempo entre o diagnóstico e o óbito ou data da última data de consulta) foi avaliada através de curvas de Kaplan-Meyer e teste de Log-Rank ou regressão de COX. Resultados: 23 mulheres com CCOC (46%), e 27 com EOC (54%) foram incluídas; 80% tinham endometriose associada, 42% eram nulíparas, 42% eram pré-menopausa e CA125 foi elevado em todos estádios (FIGO I-II= média 614.7Ui/mL; FIGO III-IV= media 2361.2Ui/mL). A média de idade ao diagnóstico foi 7 anos menor em mulheres com EOC do que naquelas com CCOC. O CCOC foi mais diagnosticado em estágios iniciais quando associado à endometriose (p=0,03). O prognóstico dos EOC e CCOC em estádios iniciais foi semelhante (p=0,96). Os CCOC não associado à endometriose tiveram menor SG (p=0,04). A expressão de todos os biomarcadores esteve presente nos EAOC e na endometriose. O aumento da expressão de VEGF entre endometriose e câncer foi significativo (p=0,0002). A hiperexpressão de PARP-1 correlacionou-se negativamente com a SLP (p=0,03) e SG (p=0,01) em estádios iniciais. Conclusão: Os CCOC e EOC são comumente diagnosticados em estádios iniciais (FIGO I-II= 68%) e estão frequentemente associados à endometriose (80% dos casos). Quando associados à endometriose, os CCOC foram mais diagnosticados em estádios iniciais e tiveram SG maior. Houve elevada porcentagem de células com ARID1A mutado nos EAOC (>40%). VEGF se expressou mais intensamente nos CCOC e EOC que na endometriose, já a expressão de PD-L1 e de PARP-1 foi similar. Apenas a hiperexpressão de PARP-1 reduziu significativamente a SLP e a SG nos CCOC e EOC nos estádios iniciais(AU)

Abstract: Introduction: Endometriosis is a benign disease, able to progress widely and generate atypical clones. It is a precursor of clear cell ovarian carcinomas (CCOC) and endometrioid ovarian carcinomas (EOCs), now called endometriosis-associated ovarian carcinomas (EAOC). Objectives: To compare the epidemiological profile, association with endometriosis and the expression of immunohistochemical markers for ARID1A, VEGF, PD-L1 and PARP-1 in women with CCOC and EOC, and its correlation with progression-free survival (PFS) and overall survival (OS). Methods: A reconstituted cohort study with 50 cases of CCOC and EOC included. Cases were treated at CAISM-UNICAMP between 1995 and 2016, followed up until 02/2017. Tissue microarrays with CCOC, EOC and endometriosis samples were stained with monoclonal antibodies against ARID1A, and for VEGF, PD-L1, PARP-1 biomarkers by immunohistochemistry. The expression of ARID1A was classified (0 to 100) according to the percentage of unstained cells. The expression of VEGF, PD-L1 and PARP-1 was classified (0 to 300) multiplying the percentage of stained cells by an intensity of expression factor (absent=0, weak=1, moderate=2, strong=3). Age at diagnosis; menopause; BMI (body mass index); CA-125 levels; diagnosis of endometriosis; date of diagnosis, date of progression, date of death and date of last consultation were retrieved from the medical records. Comparison between groups was performed through T and ?2 tests. The PFS (difference in time between diagnosis and progression date) and OS (difference in time between diagnosis and death or the last date of consultation) was assessed using Kaplan-Meyer curves and Log-Rank test or COX multivariate models. Results: twenty-three women with CCOC (46%), and 27 with EOC (54%) were included; 80% had associated endometriosis, 42% were nulliparous, 42% were premenopausal, and CA125 was elevated at all stages (FIGO I-II = mean 614.7Ui / mL; FIGO III-IV = mean 2361.2Ui / mL). The mean age at diagnosis was 7 years lower in women with EOC than in those with CCOC. CCOC when associated with endometriosis were more diagnosed at early stages (p=0.03). The prognosis of EOC and CCOC at early stages was similar (p=0.96). CCOCs not associated with endometriosis had shorter OS (p=0.04). Expression of all biomarkers was present in the EAOC and endometriosis. The increase in VEGF expression between endometriosis and cancer was significant (p=0.0002). The overexpression of PARP-1 correlated negatively with PFS (p=0.03) and OS (p=0.01) at FIGO I-II stages. Conclusion: The diagnosis of women with EOC was made earlier than in those with CCOC. CCOC and EOC are commonly diagnosed in early stages (FIGO I-II - 68%) and were associated with endometriosis (80% of cases). When associated with endometriosis, clear cell carcinomas are more likely diagnosed at early stages, and the association of endometriosis with CCOC improves OS. There was a high percentage of cells with mutated ARID1A gene in EAOC (> 40%). VEGF was expressed more intensely in CCOC and EOC than in endometriosis, whereas expression of PD-L1 and PARP-1 was similar. Only the overexpression of PARP-1 significantly reduced PFS and OS in CCOC and EOC at early stages(AU)

Antitumor activity of a novel PARP1/2 inhibitor YHP-743 / 药学学报

Poly(ADP-ribose) polymerase (PARP)-1 and PARP2 function as ADP-ribosylases involved in DNA repair. PARP1/2 is highly expressed in cancers and emerged as an attractive target for antitumor drug. In this study, we investigated the antitumor activity of a novel PARP1/2 inhibitor YHP-743 in vitro and in vivo. The results showed that YHP-743 had potent enzymatic inhibitory activity against PARP1 and PARP2 to down-regulate the PAR level. YHP-743 not only inhibited breast cancer cells with genes deficiency of homologous recombination repair, but also potentiated chemotherapy agent's cytotoxicity, such as temozolomide, topotecan, cisplatin and doxorubicin. YHP-743 elicited good antitumor activity in combination with temo-zolomide in vivo.

Effect of PARP-1 on cisplatin resistance in breast cancer MCF-7 cells / 中国病理生理杂志

AIM:To study the effect of poly(ADP-ribose)polymerase-1(PARP-1)on cisplatin resistance of human breast cancer MCF-7 cells and its possible mechanisms.METHODS: The expression of PARP-1 at mRNA and protein levels in MCF-7 cells and MCF-7/DDP cells was determined by real-time PCR and Western blot.The expression of PARP-1 in the MCF-7/DDP cells was blocked by PARP-1 siRNA.The cell viability and apoptosis were detected by the CCK-8 assay and flow cytometry analysis,respectively.Furthermore, the protein levels of PARP-1, Bcl-2, Bax, cleaved caspase-3,caspase-3,cytochrome C(Cyto-C),extracellular signal-regulated kinase(ERK)and phosphorylated ERK(p-ERK)were detected by Western blot.RESULTS:The expression of PARP-1 at both mRNA and protein levels was signifi-cantly up-regulated in the MCF-7/DDP cells.The expression of PARP-1 was increased in the MCF-7 cells treated with cis-platin.Knockdown of PARP-1 induced the apoptosis of MCF-7/DDP cells with an increased sensitivity to cisplatin.Mean-while,knockdown of PARP-1 down-regulated the protein levels of Bcl-2/Bax and p-ERK, but up-regulated the protein levels of cleaved caspase-3 and Cyto-C.After incubated with a specific ERK inhibitor U 0126,the cell viability in PARP-1 siRNA group was down-regulated significantly.CONCLUSION:Knockdown of PARP-1 increases the sensitivity of MCF-7/DDP cells to cisplatin,and promotes the cell apoptosis via mitochondrial apoptosis pathway.The mechanism may be re-lated to the attenuation of ERK signaling pathway by inhibiting phosphorylation of ERK.

Effect of nano-SiO_2 on the survival and PARP-1 expression in 16HBE cells / 中国职业医学

OBJECTIVE: To explore the effects of nano-silicon dioxide( SiO_2) on the survival and poly( ADP-ribose)polymerase-1( PARP-1) expression in human bronchial epithelial cells( 16 HBE cells). METHODS: i) The 16 HBE cells were treated with nano-SiO_2 at concentrations ranging from 0 to 100 mg/L for 24. 0 hours,and CCK-8 assay was used to examine cell viability. ii) The 16 HBE cells were divided into 6 groups: solvent control group( equal volume solvent treatment),micro-SiO_2 control group( treated with 20 mg/L micro-SiO_2),5,10,and 20 mg/L nano-SiO_2 groups( treated with the corresponding final dose of nano-SiO_2),and curcumin group. The curcumin group was given pretreatment with curcumin at a final concentration of 10 μmol/L for 2. 0 hours followed by treatment with a final concentration of 20 mg/L of nano-SiO_2. Cells in each group were harvested at time points of 4. 0,12. 0 and 24. 0 hours after treatment. The relative expression of PARP-1 mRNA and protein in 16 HBE cells was detected by quantitative real-time polymerase chain reaction and Western blotting respectively. RESULTS: i) The survival of 16 HBE cells decreased with increasing nano-SiO_2 treatment dose,showing a dose-effect relationship( P < 0. 01). ii) The expression of PARP-1 mRNA and protein in 16 HBE cells were dose-dependently decreased after nano-SiO_2 stimulation at the 12. 0 and 24. 0 hours time points( P < 0. 01). The expression of PARP-1 mRNA and protein in 5,10,and 20 mg/L nano-SiO_2 groups decreased at the above mentioned time points( P < 0. 05),compared with the solvent control group at the same time points. The expression of PARP-1 mRNA and protein in 20 mg/L nano-SiO_2 group was lower than that in the micro-SiO_2 control group at the same 12. 0 and 24. 0 hours time point( P < 0. 05). The above two indexes of cells were higher in curcumin group than that of 20 mg/L nano-SiO_2 group at the 12. 0 hours time point( P < 0. 05). CONCLUSION: Nano-SiO_2 stimulation can lead to decrease survival of 16 HBE cells in a dose-dependent manner and down-regulation of PARP-1 expression may be one of the mechanisms of proliferation and inhibition of 16 HBE cells induced by nano-SiO_2. Curcumin has certain protective effect on nano-SiO_2-induced 16 HBE cell injury.

Expression of DNA Damage Response Proteins and Associations with Clinicopathologic Characteristics in Chinese Familial Breast Cancer Patients with BRCA1/2 Mutations / 한국유방암학회지

PURPOSE: The characteristic expression of DNA damage response proteins in familial breast cancers with BRCA1, BRCA2, or non-BRCA1/2 mutations has not been analyzed in Chinese patients. Our study aimed to assess the differential expression of microcephalin 1 (BRIT1), ATM serine/threonine kinase (ATM), checkpoint kinase 2 (CHEK2), BRCA1, RAD51 recombinase (RAD51), and poly (ADP-ribose) polymerase 1 (PARP-1) and establish the profile of Chinese familial breast cancers with different mutation status. METHODS: We constructed five tissue microarrays from 183 familial breast cancer patients (31 with BRCA1 mutations; 14 with BRCA2 mutations, and 138 with non-BRCA1/2 mutations). The DNA response and repair markers used for immunohistochemistry analysis included BRIT1, ATM, CHEK2, BRCA1, RAD51, and PARP-1. The expressions of these proteins were analyzed in BRCA1/2 mutated tumors. The association between pathologic characteristics with BRCA1/2 mutation status was also analyzed. RESULTS: In familial breast cancer patients, BRCA1 mutated tumors were more frequent with high nuclear grade, estrogen receptor/progesterone receptor/human epidermal growth factor receptor 2 negative, low Ki-67, and positive CK5/6. BRCA1 mutated tumors had lower CHEK2 and higher cytoplasmic BRIT1 expression than BRCA2 and non-BRCA1/2 mutation tumors. BRCA2-associated tumors showed higher CHEK2 and cytoplasmic RAD51 expression than those in other groups. Nuclear PARP-1 expression in BRCA1/2-associated tumors was significantly higher than in non-BRCA1/2 mutation tumors. Moreover, we found quite a few of negative PARP-1 expression cases in BRCA1/2 mutated groups. CONCLUSION: The clinicopathologic findings of BRCA1-associated Chinese familial breast cancers were similar to the results of other studies. Chinese familial breast cancer patients with BRCA1/2 mutations might have distinctive expression of different DNA damage response proteins. The reduced expression of PARP-1 in Chinese BRCA1/2 mutated breast cancer patients could influence the therapeutic outcome of PARP-1 inhibitors.

Research advances in the relationship between PARP-1 and its inhibitors and the development, progression,and radiosensitivity of tumors / 中华放射肿瘤学杂志

Poly(ADP-ribose)polymerase-1(PARP-1),a ubiquitous and abundant nuclear protein composed of amino-acid residues, plays an important role in the process of DNA damage repair. In recent years,some studies have confirmed that PARP-1 is closely related to the development and progression of tumors,and may influence the tumor radiosensitivity.This article summarizes the relationship of PARP-1 and its inhibitors and the development,progression,and radiosensitivity of tumors.

Effect of silencing of poly(ADP-ribose) polymerase-1 on cell apoptosis induced by hydroquinone in rat bone marrow mesenchymal stem cells / 中国职业医学

OBJECTIVE: To explore the effect of silencing of poly( ADP-ribose) polymerase-1( PARP-1) on cell apoptosis induced by hydroquinone( HQ) in rat bone marrow-derived mesenchymal stem cells( BMMSCs). METHODS: i) The RNA expression vectors for PARP-1 gene were transfected into BMMSCs. Neomycin was used to select the transfected cells that stably expressed PARP-1-shRNA. Western blotting was used to examine the gene silencing efficiency. ii) BMMSCs with PARP-1 silencing were sorted as the treated group,while BMMSCs with empty vector were considered as the control group.HQ dissolved in phosphate buffer solution at the concentrations of 0. 0,2. 5,5. 0,10. 0,20. 0,40. 0,80. 0,160. 0 and320. 0 μmol / L were given to both groups for 24 hours. The cell viability was detected by methyl thiazolyl tetrazolium assay,and the concentration of HQ was chosen for the following study based on cell viability. iii) Both groups were treated by HQ at concentrations of 0. 0-20. 0 μmol / L for 24 hours,then the apoptosis of BMMSCs was detected by flow cytometry. The PARP-1 mRNA expression was determined by real-time fluorescent quantitative polymerase chain reaction assay. RESULTS: i) PARP-1 silencing cells and empty vector control cells were successfully screened with a mass concentration of 400 mg / L neomycin,and confirmed by level of protein expression. The interference efficiency of PARP-1 gene and inhibition efficiency was 85. 00%. ii) Based on the result of cell viability,HQ at concentrations of 0. 0-20. 0 μmol / L was chosen for the following study. iii) Compared with the group treated by HQ at concentration of 0. 0 μmol / L,the rate of early apoptosis of control group increased significantly with HQ at concentration of 10. 0 μmol / L while that of treated group was increased significantly at concentration of 5. 0 μmol / L( P < 0. 05). In addition,at concentrations of 0. 0-10. 0 μmol / L,the rates of early apoptosis in both groups increased in a dose-dependent manner( P < 0. 01). Compared with the group treated by HQ at concentrations of 0. 0 μmol / L,the expression of PARP-1 mRNA of both groups increased significantly at the concentration of 5. 0 μmol / L( P < 0. 05). The expression of PARP-1 mRNA of treated group was less than that of control group with HQ at every concentration( P < 0. 05). At concentrations of 0. 0-20. 0 μmol / L,the expression of PARP-1 mRNA of both groups increased in a dose-dependent manner( P < 0. 01). CONCLUSION: Silencing PARP-1 in BMMSCs caused cell apoptosis. PARP-1 may participate in cell apoptosis induced by HQ.

Spermidine is protective against kidney ischemia and reperfusion injury through inhibiting DNA nitration and PARP1 activation / 대한해부학회지

Kidney ischemia and reperfusion injury (IRI) is associated with a high mortality rate, which is attributed to tubular oxidative and nitrative stresses; however, an effective approach to limit IRI remains elusive. Spermidine, a naturally occurring polyamine, protects yeast cells against aging through the inhibition of oxidative stress and necrosis. In the present study, spermidine supplementation markedly attenuated histological damage and kidney dysfunction during IRI. In addition, exogenous spermidine potently inhibited poly(ADP-ribose) polymerase 1 (PARP1) activation and DNA nitrative/oxidative stress following IRI. Conversely, inhibition of ornithine decarboxylase (ODC) via siRNA transfection in vivo significantly enhanced DNA nitration, PARP1 activation, and functional damage during IRI. Finally, in ODC knockdown kidneys, PARP1 inhibition attenuated histological and functional damage induced by IRI, but not DNA nitrative stress. In conclusion, these data suggest that spermidine protects kidneys against IRI through blocking DNA nitration and PARP1 activation and this finding provides a novel target for prevention of acute kidney injury including IRI.

Expression of ERCC1, MSH2 and PARP1 in non-small cell lung cancer and their prognostic value for patients treated with platinum-based chemotherapy / 肿瘤研究与临床

Objective To analyze the expression of excision repair cross-complementation group 1 (ERCC1),MutS protein homolog 2 (MSH2) and poly ADP-ribose polymerase 1 (PARP1) in non-small cell lung cancer (NSCLC) and their prognostic value for patients receiving platinum-based postoperative adjuvant chemotherapy.Methods Immunohistochemistry was applied to detect the expression of ERCC1,MSH2 and PARP1 in 111 cases of NSCLC paraffin embedded surgical specimens.Through OG-Rank survival analysis,the prognostic value of the ERCC1,MSH2,PARP1 and the related clinic pathological factors were evaluated.Cox regression analysis was used to determine whether ERCC1,MSH2 and PARP1 were independent prognostic factors or not.Results Among the 111 NSCLC patients,the positive expression rates of ERCC1,MSH2 and RARP1 were 33.3 ％(37/111),36.9 ％(41/111) and 55.9 ％(62/111),respectively.Besides,a total of 72 patients who received platinum-based chemotherapy had complete follow-up data.ERCC1 (P＜ 0.001) and PARP1 (P=0.033) were found to be correlated with the survival time while there was no correlation for MSH2 (P =0.298).Patients with both ERCC1 and PARP1 negative had significant longer survival time than those with ERCC1 (P=0.042) or PARP1 (P=0.027) positive alone.Similarly,the survival time of patients with both ERCC1 and PARP1 positive was shorter than those with ERCC1 (P=0.048) or PARP1 (P=0.010) positive alone.Conclusions The NSCLC patients with ERCC1 and (or) PARP1 negative may benefit from platinumbased postoperative adjuvant chemotherapy.The detection of ERCC1 and PARP1 can be used as an important method to assess the prognosis of patients with NSCLC.

Role of Shh-PARP-1 signaling pathway in the protective effects of tea polyphenols against fatty acid-induced injury to islet microvessel endothelial function / 第二军医大学学报

Objective To investigate the role of Shh-PARP-1 signaling pathway in the protective effects of tea polyphenols against the fatty acid-induced islet microvessel endothelial function injury. Methods Mouse islet microvessel endothelial MS-1 cells were used in this study, and the cells were divided into normal control group, solvent group, fatty acid group (0. 25 mmol/L palmitic acid + 0. 5 mmol/L oleic acid), tea polyphenols group (25 μmol/L), fatty acid + tea polyphenols group, PARP-1 inhibitor (8 μmol/L BYK204165) + fatty acid group, PARP-1 inhibitor + fatty acid + tea polyphenols group, Shh inhibitor (2. 5 μmol/L cyclopamine) + fatty acid group, Shh inhibitor + fatty acid + tea polyphenols group and inhibitors of Shh and PARP-1 + fatty acid +tea polyphenols group. The changes of cell viability, apoptosis, nitric oxide (NO) synthesis and oxidative stress related indicators were examined in each group. Results After fatty acid treatment, the survival rate ofMS-1 cells was decreased, and the level of apoptosis was significantly increased (P0. 05). Conclusion Fatty acid can directly trigger islet microvessel endothelial function injury, and tea polyphenols shows a protective effect against the toxicity of fatty acid, which can be enhanced by inhibiting Shh-PARP-1 signal pathway.

Role of hydrogen gas in regulating of poly (ADP-ribose) polymerase-1 dependent cell death in rat Schwann cells / 中华危重病急救医学

Objective To investigate the protective effects and underlying molecular mechanisms of hydrogen (H2) on high glucose-induced poly (ADP-ribose) polymerase-1 (PARP-1) dependent cell death (PARthanatos) in primary rat Schwann cells. Methods Cultured primary rat Schwann cells were randomly divided into five groups: blank control group (C group), H2 control group (H2 group), high osmotic control group (M group), high glucose treatment group (HG group), and H2 treatment group (HG+H2 group). The cells in H2 group and HG+H2 group were cultured with saturated hydrogen-rich medium containing 0.6 mmol/L of H2, and those in three control groups were cultured with low sugar DMEM medium containing 5.6 mmol/L of sugar, and the cells in HG and HG+H2 groups were given 44.4 mmol/L of glucose in addition (the medium containing 50 mmol/L of glucose), the cells in C group and H2 group were given the same volume of normal saline, and the cells in M group were given the same volume of mannitol. Cytotoxicity was evaluated using lactate dehydrogenase (LDH) release rate assays after treatment for 48 hours in each group. The contents of peroxynitrite (ONOO-) and 8-hydroxy-2-deoxyguanosine (8-OHdG) reflecting oxidative stress injury and DNA damage were detected by enzyme linked immunosorbent assay (ELISA). Poly (ADP-ribose) (PAR) protein expression was analyzed by Western Blot, and immunofluorescence staining was used to determine the nuclear translocation of the apoptosis-inducing factor (AIF). Results The cytotoxicity in HG and HG+H2 groups was significantly increased as compared with that of C group [LDH release rate: (61.40±2.89)%, (42.80±2.32)% vs. (9.92±0.38)%, both P < 0.01], the levels of ONOO- and 8-OHdG were markedly elevated [ONOO- (ng/L): 853.58±51.00, 553.11±38.66 vs. 113.56±14.22; 8-OHdG (ng/L): 1 177.37±60.97, 732.06±54.29 vs. 419.67±28.77, all P < 0.01], and the PAR protein expression was up-regulated (A value: 0.603±0.028, 0.441±0.010 vs. 0.324±0.021, both P < 0.01). The cytotoxicity, the levels of ONOO- and 8-OHdG, and PAR expression in HG+H2 group were significantly lower than those of the HG group (all P < 0.01). There were no significant differences in above parameters between H2 group as well as M group and C group. It was shown by immunofluorescence that AIF was expressed in the cytoplasm in C group, H2 group and M group, AIF was expressed in the whole cell in HG group, and the expression in the nucleus was particularly increased. A small amount of AIF expression was found in the nucleus of HG+H2 group, which indicated that high glucose could promote the AIF nuclear translocation, and that hydrogen-rich medium could prevent the process of translocation. Conclusions High glucose levels could enhance DNA damage that enhance PARthanatos in primary rat Schwann cells. However, H2 can not only reduce DNA damage of injured cells, but also inhibit the special death process, reduce the cell toxicity, all of which have protective effects.

Expression of PARP-1 in epithelial ovarian cancer and its relationship with epithelial-mesenchymal transition / 中国病理生理杂志

AIM:To investigate the expression of poly (ADP-ribose) polymerase-1 (PARP-1) in the epithelial ovarian cancer ( EOC) and its relationship with epithelial-mesenchymal transition ( EMT) .METHODS:The expression of PARP-1, E-cadherin, vimentin and Snail was detected in the EOC and benign ovarian tumor tissues by immunohistochemi -cal method and real-time PCR.The expression of PARP-1, E-cadherin, vimentin and Snail proteins in the SKOV3 cells treated with efficient PARP-1 inhibitor PJ34 was determined by Western blotting .RESULTS:The positive expression rates of PARP-1, vimentin and Snail were significantly higher in the EOC than that in the benign ovarian tumor tissues , whereas the positive expression rate of E-cadherin was the opposite (P<0.05).The expression of PARP-1, E-cadherin, vimentin and Snail in the EOC was associated with the histological grade , clinical stage and lymphatic metastasis (P<0.05), but no relationship with age and pathological types was observed .The expression of E-cadherin in the EOC was negatively co-related to that of PARP-1.In contrast, the expression of vimentin and Snail in the EOC was positively co-related to that of PARP-1.The relative mRNA expression of PARP-1, vimentin and Snail in the EOC was significantly higher than that in the benign ovarian tumor tissues ( P<0.05) , while the mRNA expression of E-cadherin in the EOC was remarkably lower than that in the benign ovarian tumor tissues (P<0.05).The protein expression of PARP-1, vimentin and Snail in the SKOV3 cells was significantly decreased (P<0.05), while E-cadherin protein was increased after treated with PJ 34(P<0.05). CONCLUSION:PARP-1 may contribute to the onset of EMT in the EOC by regulating the expression of E -cadherin, vim-entin and Snail.The role of PARP-1, which is relevant to EMT, might be important in the development of ovarian cancer .

Influence of Poly (ADP-ribose) Polymerase Inhibitor PJ34 on Blood Brain Barrier in Rats with Cerebral Ischemia-reperfusion Injury / 中国医科大学学报

Objective To investigate the influence of poly(ADP-ribose)polymerase inhibitor PJ34 on blood brain barrier(BBB)in rats with cere-bral ischemia-reperfusion injury. Methods Rat model of cerebral ischemia-reperfusion injury was established by the middle cerebral artery occlu-sion. A total of 135 SD rats were randomly divided into 3 groups:sham-operated group(sham group),ischemia-reperfusion group(IR group)and PJ34 group(PJ34 group). 45 animals in each group were then equally divided into subgroups and the rats were sacrificed at 6 h,24 h,48 h after re-perfusion,respectively. BBB permeability was evaluated by detection of extravasated Evans blue(EB). The expression of tumor necrosis factorα(TNF-α)and matrix metalloproteinase 9(MMP-9)activity were measured by immunohistochemistry and western blot at different time points. Re?sults Compared with sham group,the contents of EB and the expressions of TNF-αand MMP-9 in IR group were increased significantly(P<0.05). Compared with IR group,the contents of EB and the expressions of TNF-αand MMP-9 in PJ34 group were markedly decreased at the same time point(P<0.05). Conclusion The present study provided in vivo evidence that PARP inhibitor PJ34 can protect against cerebral ischemia re-perfusion injury,and the mechanism might be related to maintaining the stability of blood-brain barrier by suppressing the expression of TNF-αand MMP-9 in ischemic cortex.

Cisplatin induces primary necrosis through poly(ADP-ribose) polymerase 1 activation in kidney proximal tubular cells / 대한해부학회지

Treatment with cisplatin for cancer therapy has a major side effect such as nephrotoxicity; however, the role of poly (ADP-ribose) polymerase 1 (PARP1) in necrosis in response to cisplatin nephrotoxicity remains to be defined. Here we report that cisplatin induces primary necrosis through PARP1 activation in kidney proximal tubular cells derived from human, pig and mouse. Treatment with high dose of cisplatin for 4 and 8 hours induced primary necrosis, as represented by the percentage of propidium iodide-positive cells and lactate dehydrogenase release. The primary necrosis was correlated with PARP1 activation during cisplatin injury. Treatment with PJ34, a potent PARP1 inhibitor, at 2 hours after injury attenuated primary necrosis after 8 hours of cisplatin injury as well as PARP1 activation. PARP1 inhibition also reduced the release of lactate dehydrogenase and high mobility group box protein 1 from kidney proximal tubular cells at 8 hours after cisplatin injury. Oxidative stress was increased by treatment with cisplatin for 8 hours as shown by 8-hydroxy-2'-deoxyguanosine and lipid hydroperoxide assays, but PARP1 inhibition at 2 hours after injury reduced the oxidative damage. These data demonstrate that cisplatin-induced PARP1 activation contributes to primary necrosis through oxidative stress in kidney proximal tubular cells, resulting in the induction of cisplatin nephrotoxicity and inflammation.

Expression of Poly ADP-Ribose Polymerase-1 in Human Breast Cancer Tissue and Its Relationship with Prognosis Related Factors / 中国康复理论与实践

@#Objective To investigate the expression of poly ADP-ribose polymerase-1(PARP-1) in human breast cancer tissue and its relationship with patient's prognosis related factors.Methods The expression of PARP-1 in breast cancer tissues and their adjacent non-cancerous tissues from resection in 66 operated cases were detected using RT-PCR and immunofluorecence staining(IFS).The relationship between the expression of PARP-1 and the prognosis related factors was analyzed.Results Compared with that of the adjacent non-cancerous tissues,the expression of PARP-1 mRNA increased in 94.9%(62/66) patients,while the expression of PARP-1 protein increased in 90.9%(60/66) ones.The label index(LI) and the expression index(EI) of PARP-1 gene increased especially in the cases in later clinical stage,with poor tumor differentiation grade,of pure breast carcinomas or infiltrative breast carcinomas and of metastasis.Conclusion The expression of PARP-1 in human breast cancer tissue on the level of transcription and translation increases,which may be positively related with the patient's prognosis.

The Role of Poly(ADP-ribose) Polymerase-1 in Ventilator-Induced Lung Injury / 결핵및호흡기질환

BACKGROUND: Reactive oxygen species (ROS) take center stage as executers in ventilator-induced lung injury (VILI). The protein with DNA-damage scanning activity, poly (ADP-ribose) polymerase-1 (PARP1), signals DNA rupture and participates in base-excision repair. Paradoxically,overactivation of PARP1 in response to massive genotoxic injury such as ROS can induce cell death through beta-nicotinamide adenine dinucleotide (NAD+) depletion, resulting in inflammation. The purpose of this study is to investigate the role of PARP1 and the effect of its inhibitor in VILI. METHODS: Forty-eight male C57BL/6 mice were divided into sham, lung protective ventilation(LPV), VILI, and PARP1 inhibitor (PJ34)+VILI (PJ34+VILI) groups. Mechanical ventilator setting for the LPV group was PIP 15 cmH2O + PEEP 3 cmH2O + RR 90/min + 2 hours. The VILI and PJ34+VILI groups were ventilated on a setting of PIP 40 cmH2O + PEEP 0 cmH2O + RR 90/min + 2 hours. As a PARP1 inhibitor for the PJ34+VILI group, 20 mg/Kg of PJ34 was treated intraperitoneally 2 hours before mechanical ventilation. Wet-to-dry weight ratio and acute lung injury (ALI) score were measured to determine the degree of VILI. PARP1 activity was evaluated by using an immunohistochemical method utilizing biotinylated NAD. Myeloperoxidase (MPO) activity and the concentration of inflammatory cytokines such as tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and IL-6 were measured in bronchoalveolar lavage fluid (BALF). RESULTS: In the PJ34+VILI group, PJ34 pretreatment significantly reduced the degree of lung injury, compared with the VILI group (p<0.05). The number of cells expressing PARP1 activity was significantly increased in the VILI group, but significantly decreased in the PJ34+VILI group (p=0.001). In BALF, MPO activity, TNF-alpha, IL-1beta, and IL-6 were also significantly lower in the PJ34+VILI group (all, p<0.05). CONCLUSION: PARP1 overactivation plays a major role in the mechanism of VILI. PARP1 inhibitor prevents VILI, and decreases MPO activity and inflammatory cytokines.