| Objective: Ovarian cancer is one of the three common malignancies thatfound in the female genital system. Because of its deep location in the pelvisand asymptomatic nature, Ovarian cancer is characterized by presentation atan advanced stage, so ovarian cancer mortality rate ranks first in gynecologicmalignancies, has become a serious threat in the primary tumor to women’slives and health. Epithelial ovarian cancer accounts for nearly80%of ovarianmalignant tumors. The incidence of occult, early asymptomatic, oftendiscovered at an advanced stage, which make the early diagnosis and earlytreatment extremely difficult.For the treatment of ovarian cancer, appropriate and effective surgerycombined with chemotherapy can significantly improve patient survival.Epithelial ovarian cancer is very sensitive to chemotherapy drugs. Thetraditional chemotherapy has been the use of paclitaxel plus platinum, butmost patients relapse after stopping the medication for some time, this maycaused by the unclear pathogenesis of ovarian cancer and the emergence oftumor drug resistance.Bradykinin (BK), a vasoactive peptide released during inflammation, isthought to be a multi-potential stimulant of cancer growth. BK exerts itscellular effects through binding two pharmacologically distinct G-protein-coupled receptors (GPCRs), the B1receptor(B1R) and the B2receptor(B2R) located on the cell membrane.B2R is constitutively expressed in most tissues where it involves avariety of physiological and biochemical processes of the body; B1R is aninducible kinin receptor that becomes expressed and active only afterinflammation injury or cytokine stimulation. High BK concnentrations werefound in ascitic and pleural fluids obtained from patients with gastric,ovarian and hepatic cancers. Moreover, evidence indicates that major cellularelements of the tumer microenvironment express either B1R or B2R or bothand their stimulation modulates cell prolifecation, angiogenesis of the tumercells. However, But the reports on the expression of B1R and B2R in ovariancancer cells are not consistent and rare.At present, BK receptor antagonists have been tested in a variety oftumors, especially the treatment for lung cancer and prostate cancer, but itstrial in ovarian cancer research is less. Here, we carried on the comparativestudy on the expression of B1R, B2R, eNOS and iNOS in ovarian tissuesderived from epithelial ovarian cancer and ovarian benign tumor patients aftersurgical removal of the tumor tissue. On this basis, we observed the inhibitoryeffects of B2receptor antagonist HOE-140alone or in combination withdoxorubicin (DOX) on human ovarian cancer cell lines (SKOV3) andpossible mechanisms of action. The study is expected to find a better targetfor the tumor, thereby improving the quality of life of patients with ovariancancer.Method:1Expression of B1R, B2R, eEOS and iNOS in ovarian cancer tissue.20patients with histologically confirmed epithelial ovarian cancer andAnother20patients with ovarian benign tumors were identified from a tissue,ages ranges from35to55years old, without hypertension, diabetes and othermedical problems and a history of genetic disease.The expressions of B1R, B2R, eEOS and iNOS in ovarian cancer tissue weredetected by immunohistochemistry after the slices were pathologicallydiagnosed as benign ovarian tumors or ovarian cancer.2Determination of serum Nitric Oxide (NO) levelSerum was collected from the patients diagnosed as ovarian cancerpatients and healthy volunteers prior to definitive therapy, and serum NOlevels were determined by using a NO detection kit.3Cytotoxic effect of HOE-140in the ovairan cancer SKOV3cells-synergistic interaction between HOE-140and DOX. SKOV3cells were cultured in the absence (vehicle control) or in thepresence of increasing concentrations of HOE-140(0.01,0.1,1,10and100μmol/L) or DOX (3,10,30,100and300μmol/L) for24h. Then, thecombination of HOE-14010μmol/L and DOX10μmol/L were used toobserve the synergistic interaction.4DOX and HOE-140alone or in combination on apoptosis of SKOV3cellsThe SKOV3cells were treated with DOX10μmol/L, HOE-14010μmol/L or a combination of DOX10μmol/L+HOE-14010μmol/L for24h, apoptosis of SKOV3cells induced by DOX or HOE-140was examined byThe terminal deoxynucleotidyltransferase-mediated dUTP-biotin nickend-labeling (TUNEL) assay.Results:1Expression of B1R, B2ER, eNOS and iNOS in ovarian tissue samplesOvarian tissue from the patients diagnosed with ovarian cancer or benignovarian disease were used to detect the expression of B1R, B2R, eNOS andiNOS by immunohistochemistry. The results showed that almost no B1R andiNOS expressions were seen in benign ovarian tissue, while there were someexpressions of B2R and eNOS in benign ovarian tissue. Compared withbenign ovarian tissue, high expression of B1R, B2R, eNOS and iNOS wereseen in all ovarian cancers.2Serum concentrations of NOSerum No levels were obviously higher in patients with ovarian cancerthan that in healthy volunters(P<0.01).3Cytotoxic effect of HOE-140in the ovairan cancer SKOV3cells-synergistic interaction between HOE-140and DOX.MTT proliferation assay results showed that both HOE-140and DOXinhibited the proliferation of SKOV3cells in a concentration-dependentmanner。The inhibition ratios of HOE-140and DOX at10μmol/L were12.7%and26.8%respectively, which were smaller than that of thecombination effects (37.5%). The effect of HOE-140and DOX combination on SKOV3cells showed a synergistic effects (P <0.01).4DOX and HOE-140alone or in combination on apoptosis of SKOV3cellsCell morphology of SKOV3cells by TUNEL staining assay (on24h)showed Cytoplasmic condensation and diverse morphology in the presence ofHOE-14010μmol/L or DOX10μmol/L, the combination effect of the twodrug was stronger than medication alone.Conclusions1High serum NO levels were found in patients with ovarian cancer, alsohigh expressions of B1R, B2R, eNOS and iNOS were found in ovarian cancertissues;2HOE-140alone or combination with DOX inhibited SKOV3cell Prol-ifer ation, its mechanism may be caused by inducing cell apoptosis andinhibiting NO production. |
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