treatment with 300 μg / ml ole downregulated invasion - promoting mir - 181b - 3p, mir - 221 - 3p, radiosensitivity - promoting mir - 16 - 5p, and anti - apoptotic bcl - 2 and mcl1, while upregulating invasion - inhibiting mir - 29a - 3p, mir - 34a - 5p, mir - 125 - 5p and apoptosis - promoter fas, p53, tnf receptor superfamily member 10b ( tnfrsf10b9, also known as trail ), and bid. ovarian cancer is a highly aggressive neoplasm whose complex management ( due to molecular and cellular heterogeneity ) together with its commonly delayed diag nosis have made it the fifth cause of death in women. the great majority of ovarian cancers are classified as epithelial belonging to the serous subtype. treatment of high - grade ovarian serous adenocarcinoma cell line hey with 200 μm and 400 μm ole led to a reduction in cell viability, with accumulation of cells in g2 / m phase and induction of apoptosis. a detailed analysis revealed that 400 μm ole acted as a pro - oxidant agent, increasing oxygen radical generator labile iron pool ( lip ) and ros levels. similarly, treatment of ovarian serous adenocarcinoma ovcar - 3 cell line with 200 μg / ml olive leaf extract containing 87 % ole for 24 h induced s / g2m phase cell cycle arrest and apoptosis. these effects were dependent on ole - mediated ros accumulation and reduction in cat and sod2 protein levels. prostate cancer is the fifth leading cause of cancer - related deaths among men, whose therapeutic options are mainly defined on the basis of a combination of life expectancy, disease stage / risk classification / presence of metastasis, and cancer sensitivity towards androgens. treatment of benign prostatic hyperplasia ( bph ) epithelial cell line bph - 1 and prostate cancer cell lines lncap and du145 with 100 μm and 500 μm ole for 72 h determined a reduction in cell viability, probably as a consequence of the reduction in akt phosphorylation levels, with cells showing signs of necrosis for the highest of the two tested ole doses. treatment with 100 μm and 500 μm ole for 72 h also produced different effects on ros homeostasis in the three cell lines. in bph - 1, a reduction in ros was measured, while no effect was recorded for lncap cells, and a pro - oxidant activity with a marked ros increase was exerted by ole on du145 cells. measurement of non - protein thiol groups as a marker of oxidative stress defense revealed that ole caused a dose - dependent increase in thiol groups in both bph - 1 and lncap cells, but only 100 μm ole augmented thiol groups in du145 cells; on the contrary, 500 μm ole diminished thiol groups in this cell line. as regards oxidative damage protecting enzyme heme oxygenase - 1 ( ho - 1 ), both 100 μm and 500 μm ole increased ho - 1 protein levels in bhp - 1 and lncap cells, while in du145 cells, ole treatment decreased ho - 1 enzyme expression. the levels of γ - glutamylcysteine synthetase ( γ - gcs ) ( involved in reduced glutathione synthesis ) were not affected by ole treatment in the three studied cell lines. ros homeostasis also seems to play a fundamental role in ht cytotoxic action on prostate cancer cells. treatment of prostate cancer cell line pc3 with 80 μm ht caused mitochondrial dysfunction and triggered o2 - dependent apoptosis. similarly, 50 – 150 μm ht reduced cell viability of pc3 and lncap cells after 72 h treatment in a dose - dependent fashion ( mean ic50 ± s. d. = 103 ± 7 μm and 146 ± 12 μm for pc3 and lncap cells, respectively ), with a mechanism that seemed dependent on the ability of culture conditions to favor h2o2 accumulation ( quantified in the medium after 24 h incubation with 100 μm ht as mean ± s. d. = 12. 0 ± 2. 9 and 8. 8 ± 3. 4 for pc3 and lncap cells, respectively ). ht action might overcome androgen sensitivity. incubation of androgen - sensitive lncap and castration - resistant c4 - 2 cell lines with 50 – 400 μm ht for 48 and 72 h reduced cell viability in a dose - dependent manner, with the ic50 values for lncap = 190 μm and 86. 9 μm after 48 and 72 h, respectively, and the ic50 values measured for c4 – 2 = 176 μm and 76. 5 μm after 48 and 72 h, respectively. cell cycle analysis after 48 h incubation with 100 – 300 μm demonstrated a dose - dependent cellular accumulation in the g1 phase and apoptosis induction, with ( i ) caspase - 3 / 7 and parp activation, ( ii ) reduction in bcl - 2, bcl - xl, cell cycle progression drivers cyclins d1 and e, and cdk2 and cdk4 protein levels, ( iii ) increased bax protein levels, ( iv ) reduced nf - κb p65 nuclear localization, ( v ) reduction in activation levels of cell growth regulators akt and stat3, and ( vi ) decreased androgen receptor protein levels and activity. the 48 h incubation of human prostate cancer cell lines lncap, 22rv1, and pc - 3 with 30 – 300 μm ht decreased cell viability in a dose - dependent fashion. as indicated by the authors, mean ic50 ± s. e. m. = 41. 17 ± 2. 79 μm for lncap cells, 9. 32 ± 0. 50 for 22rv1 cell line, and 28. 88 ± 2. 25 for pc - 3 cells. treatment of pc - 3 cells with 30 μm and 100 μm ht for 24 h reduced the migration rate in a dose - dependent manner. treatment of 22rv1 cells with 10 μm ht reduced prostatosphere number and size after 10 days, and phosphorylation of erk1 / 2, camp response element - binding ( creb ) protein, and jnk after 24 h. osteosarcoma is a primary malignant bone tumor historically known for its extremely low survival rates even in the case of surgical resection / amputation, which have been improved by the introduction of neoadjuvant and adjuvant chemotherap y. data about ole cytotoxic effect in vitro are conflicting, and partially dependent on the used cell line and the duration of the treatment. in human 143b osteosarcoma cells, ole showed a dose - dependent inhibition of proliferation when used at concentrations of 62. 5 μm, 125 μm, and 250 μm for 24 h, and 1 μm – 250 μm for 48 h. also, 100 μm ole induced autophagy after 48 h and exhibited an anti - migratory effect after 60 h incubation. in human mg - 63 cells, 3 – 50 μg / ml ( ≈5 – 92. 5 μm ) ole for 24 h reduced cell viability in a dose - dependent manner with ic50 = 22 μg / ml ± 3. 6. another report documented no effect on mg - 63 cell viability at ole concentrations of 50 μm and 100 μm, while a dose - dependent cytotoxic effect was documented for 200 μm and 400 μm ole after 24, 48, and 72 h incubation, with ic50 = 346 μm for 48 h incubation. treatment of mg - 63 cells with 20 μg / ml ( ≈37 μm ) ole for 48 h boosted the transcription of autophagy - related genes unc - 51, like autophagy activating kinase 1 ( ulk1 ), activating molecule in becn1 - regulated autophagy protein 1 ( ambra1 ), and bcl - 2 interacting protein 3 like ( bnip3l ), as well as protein levels of ubiquitin binding protein p62, suggesting that ole activity relies on autophagy induction. in saos2 osteosarcoma cell line, 50 μm ole caused no changes in cell viability after 24, 48, and 72 h incubation. treatment with 100 μm ole affected cell viability only after 48 h incubation, causing no detectable effects after 24 and 72 h incubation. instead, 200 μm ole started being effective as a cytotoxic agent after 48 h incubation, with effects confirmed after 72 h incubation. on the contrary, 400 μm showed a consistent cytotoxic action at all the assayed time points ( 24, 48, and 72 h ). neuroblastomas are clinically heterogeneous embryonal neuroendocrine tumors, driving important research efforts towards the improvement of survival rates for patients with high - risk disease. both ole and ht exhibit a cytotoxic action against human neuroblastoma cell line sh - sy5y. after 48 h treatment, ic50 for ole was 350 μm. ole induced cell cycle arrest and apoptosis through upregulation of cdk inhibitors p53, p21, p15, and p16, together with downregulation of cyclins d1, d2, and d3, and cdk4 and cdk6.