ABSTRACT
Objective: cerebrospinal fluid [CSF] plays an important role in cortical development during the fetal stages. Embryonic CSF [E-CSF] consists of numerous neurotrophic and growth factors that regulate neurogenesis, differentiation, and proliferation. Mesenchymal stem cells [MSCs] are multi-potential stem cells that can differentiate into mesenchymal and non-mesenchymal cells, including neural cells. This study evaluates the prenatal and postnatal effects of CSF on proliferation and neural differentiation of bone marrow MSCs [BM-MSCs] at gestational ages E19, E20, and the first day after birth [P1]
Materials and Methods: in this experimental study, we confirmed the mesenchymal nature of BM-MSCs according to their adherence properties and surface markers [CD44, CD73 and CD45]. The multi-potential characteristics of BMMSCs were verified by assessments of the osteogenic and adipogenic potentials of these cells. Under appropriate in vitro conditions, the BM-MSCs cultures were incubated with and without additional pre- and postnatal CSF. The MTT assay was used to quantify cellular proliferation and viability. Immunocytochemistry was used to study the expression of MAP-2 and beta-III tubulin in the BM-MSCs. We used ImageJ software to measure the length of the neurites in the cultured cells
Results: BM-MSCs differentiated into neuronal cell types when exposed to basic fibroblast growth factor [b-FGF]. Viability and proliferation of the BM-MSCs conditioned with E19, E20, and P1 CSF increased compared to the control group. We observed significantly elevated neural differentiation of the BM-MSCS cultured in the CSF-supplemented medium from E19 compared to cultures conditioned with E20 and P1 CSF group
Conclusion: the results have confirmed that E19, E20, and P1 CSF could induce proliferation and differentiation of BM-MSCs though they are age dependent factors. The presented data support a significant, conductive role of CSF components in neuronal survival, proliferation, and differentiation
ABSTRACT
Objective: Methotrexate [MTX] is an antimetabolite drug commonly prescribed for the various cancers and autoimmune diseases. Despite its considerable therapeutic effects, nephrotoxicity is the most important side-effect of treatment with MTX. Aquaporin1 [AQP1] is a water channel proteins which is present in mammalian kidney. Raspberry fruit with antioxidant properties is able to protect biological systems from the harmful effects of free radicals. The purpose of this study was to investigate the effect of raspberry extract on expression of AQP1 and the MTX-induced nephrotoxicity in rats
Materials and Methods: In this experimental study, 60 adult male Wistar rats were divided into nine groups including control, sham, MTX treated group [single dose of 20 mg/kg of body weight [BW] MTX at the third day], raspberry treated groups [intraperitoneal [I.P] injection of 100, 200, 400 mg/kg of BW raspberry extract for ten consecutive days], MTX and raspberry treated groups. At day 11, rats were sacrificed via chloroform inhalation and kidney tissues were fixed in formalin solution for histological and immunohistochemistry analysis. The serological assays for urea, creatinine, uric acid and interleukin-6 [IL-6] levels were also performed
Results: MTX elevated serum level of the urea, creatinine, uric acid, IL-6, renal tissue damage and decreased the AQP1 expression level. Raspberry fruit extract improved the kidney function and reduced side effects of MTX in treated rats. Expression of AQP1, in a dose dependent manner was also ameliorated, as compared to control group
Conclusion: According to the findings of this study, it can be concluded that biological activity of compounds presented in raspberry fruit extract especially anthocyanins may have chemo-protective effect on kidney function and AQP1 expression in rats treated by MTX
ABSTRACT
Ophiocoma erinaceus Muller and Troschel [Ophiocomidae] is part of the extensive group of echinoderm that contains bioactive metabolites. As the anti cancer potential of brittle star saponin has not been reported against cervical cancer, the present study was conducted to evaluate the anticancer effect of extracted crude saponin. Saponin extraction was conducted using conventional method such as froth test, TLC, FTIR and erythrolysis assay. The Hela-S3 cervical carcinoma and HNCF-PI52 normal cells were treated with different concentrations of saponin fraction for 24 and 48 h. The cytotoxicity was examined by MTT, DAPI, AO/PI, Annexin V-FITC and flow cytometry. In addition, the apoptotic induced pathway was studied using caspase assay, evaluation of ROS generation and Bcl-2 mRNA level. Crude saponin showed cytotoxic properties in Hela-S3 cells [IC[50] of 23.4 micro g/mL] without significant impact against normal cells. In addition, the crude saponin increased sub-G1 peak in flow cytometry histogram of treated cells, ROS generation and caspase-3 and -9 activity [IC[50] of 11.10, 11.27 micro g/mL]. The dose dependent down regulation of Bcl-2 in treated cells demonstrated that saponin fraction can trigger intrinsic apoptotic pathway in cancer cells. This study provides valuable information about the apoptotic inducing effect of saponin fraction, which can offer new insights into the anticancer potential of saponin as a promising candidate against human cervical carcinoma
ABSTRACT
Background: Having low-grade chronic inflammation such as elevated C-reactive protein [CRP], interleukin-6 [IL-6] and tumor necrosis factor-alpha [TNF- alpha] plays a crucial role in polycystic ovary syndrome [PCOS]. This study aimed at investigating the therapeutic effects of curcumin on IL-6, CRP and TNF- alpha and symptoms of polycystic ovary syndrome
Methods: In this research, 72 female adult Wistar rats were divided into control [n=12], PCOS [n=12] and curcumin-treated PCOS groups [n=48]. PCOS was induced by injection of estradiol valerate [2mg/kg - one-step]. PCOS rats were divided into control and experimental groups which received daily intraperitoneal injection of curcumin. After 60 days of syndrome induction, ovaries were collected for histological and immunohistochemical evaluations. Serum IL-6 and CRP was detected by the ELISA kit. Data were analyzed using In-Stat 3 via one-way analysis of variance [ANOVA] and p<0.05 was considered statistically significant
Results: Histological studies showed a significant reduction in thickness of theca layer and increase in the number of corpus luteum [CL] diameter in the curcumintreated group compared with the PCOS group; also inflammatory markers such as IL-6 and CRP significantly decreased in groups treated with curcumin compared with PCOS groups. Regarding immunohistochemical analysis, the expression of TNF- alpha in granulosa layer and follicular fluid of follicles and ovarian cysts in PCOS group was more than the control group's expression. However, expression of this factor in the ovaries treated with curcumin was decreased
Conclusion: This study showed that the anti-inflammatory and antioxidant effects of curcumin on PCOS may be due to its inhibitory effect on expression and levels of TNF- alpha, serum IL-6 and CRP
ABSTRACT
Objective: Curcumin protects the liver against injury and fibrosis through suppressing hepatic inflammation, attenuating hepatic oxidative stress [OS], and inhibiting hepatic stellate cells [HSCs] activation. Non-alcoholic fatty liver disease [NAFLD] and polycystic ovary syndrome [PCOS] are considered as common metabolic disorders. Low-grade chronic inflammation with different markers, such as elevated C-reactive protein [CRP] and interleukin-6 [IL-6] levels, play a crucial role in PCOS. This study aimed to evaluate the therapeutic effects of curcumin on IL-6 and CRP levels as well as insulin resistance [IR] index on liver function in PCOS rats
Materials and Methods: In this experimental study, 90 adult Wistar rats were divided into control [n=18], sham [n=18], PCOS [n=18] and curcumin-treated PCOS groups [n=36]. PCOS group was injected subcutaneously with 2 mg estradio-valerate [E2V]. After 60 days, PCOS group was treated with curcumin [100 and 300 mg/kg body weight [BW]] for 14 days and anesthetized by chloroform. Blood and liver samples were collected for histological and serological analyses. Data were analyzed using In-Stat 3 via one-way analysis of variance [ANOVA]
Results: Histological and serological analyses showed a reduction in number of necrotic cells, IR index, as well as IL-6 and CRP levels in PCOS rats that were treated with various concentrations of curcumin
Conclusion: In this study, curcumin decreased liver inflammation by induction of insulin sensitivity and reduction of hepatic necrosis. Therefore, curcumin may be considered as protective factor against inflammatory state of PCOS
ABSTRACT
Polycystic ovary syndrome [PCOS] is the most common cause of infertility and endocrinopathy in women due to lack of ovulation. Silymarin has anti-inflammatory properties and is a modulator of the immune system. In this experimental study, the effect of silymarinon PCOS- induced rats was studied. In this experimental study, 144 adult Wistar rats were divided into groups of control, sham, PCOS [S.C. injection of 2mg estradiol valerate/ rat, once] and treated with silymarin. After induction of PCOS during 60 days, 20, 50, 100, 200 and 300 mg/kg BW silymarin was injected intraperitoneally for 10 days. Sham and control groups received DMSO and no injection, respectively. All groups were anesthetized and the serum and ovary of groups were collected in order to investigate the histological and serologic changes. Data were tested using ANOVA in Instat software and P< 0.05 was considered significant. PCOs induction resulted in elevation of follicular cyst formation, abnormal follicular development and increasing of androgen compared with control rats. Silymarin attenuated ovarian cysts and thickness of theca layer and improved hormonal balance and follicle development in PCOS rats. Silymarin can reduce the histological and hormonal symptoms of PCOS, and has a protective effect on ovary. It seems that this potency is due to the antioxidant and anti-inflammatory effects of silymarin, which can reduce cyst counts and improve the development of ovarian follicles
ABSTRACT
Improvements in cancer treatment have allowed more young women to survive. However, many cancer patients suffer from ovarian failure. Cryopreservation is one of the solutions for fertility restoration in these patients. The cryopreservation of isolated follicles is a more attractive approach in the long term. Many endocrine and paracrine factors can stimulate the granulosa cells of preantral follicles to proliferate. Melatonin acts as direct free radical scavenger and indirect antioxidant. In this study, we investigated the direct effects of melatonin on follicle development and oocyte maturation by exposing in vitro cultured mouse vitrified-warmed ovarian follicles to melatonin. In an experimental study, preantral follicles with diameter of 150-180 micro m were isolated from prepubertal mouse ovaries. Follicles were vitrified and thawed using cryolock method. They were then cultured individually for 7 days in droplets supplemented with 0, 10 and 100 pM melatonin, while ovulation was induced using epidermal growth factor [EGF] and human chorionic gonadotropin [hCG]. The survival rate of follicles and nuclear maturation of ovulated oocytes were determined. At the end of culture, significant increases in follicle survival [p<0.001] and in diameter [p<0.05] were noticed in 10 pM melatonin group compared to control group. In the 100 pM group, survival rate was not affected by melatonin. It was revealed that after induction of ovulation, total number of metaphase II oocytes in treatment groups were not influenced by melatonin [p>0.05]. Culture of mouse vitrified-warmed preantral follicles in a medium supplemented with 10 pM melatonin increased the number of surviving follicles
Subject(s)
Animals, Laboratory , In Vitro Techniques , Culture Media , Melatonin , Mice , In Vitro Oocyte Maturation TechniquesABSTRACT
Background : Although honeybee venom (BV) has been reported to induce apoptosis in different types of cancerous cells, its synergistic effects with customary anti-cancer drugs remain largely unknown. In the present study, we evaluated the cytotoxic effect of BV alone (as a natural product) and the synergistic cytological effects of this component in combination with [Pd (bpy) (Pi-Pydtc)]NO3 - a novel palladium complex on human T-cell lymphoblastic leukemia cells. To investigate the cytotoxic effect of the BV alone and in combination with palladium complex on MOLT-4 cells MTT assay was performed. In order to determine the apoptotic effects of BV separately and in combination with Pd (II) complex on these cells and its ability to induce apoptosis, morphological examination, flowcytometric analysis and caspase-3 colorimetric assay were done. Results : We found that BV induced morphological changes, namely nuclear shrinkage, and inhibited MOLT-4 cell proliferation; both effects were dose- and time-dependent. Flow cytometry by Annexin-V antibody demonstrated that BV induced apoptosis in MOLT-4 cells. Furthermore, BV induced apoptosis independently of caspase-3 in these cells. In addition, we proved a clear synergistic effect of BV on [Pd (bpy) (Pi-Pydtc)]NO3. The apoptotic pathway activated by BV in combination with Pd complex was caspase-3-dependent. Conclusions : These observations provide an explanation for the anti-proliferative properties of BV, and suggest that this agent may be useful for treating lymphoblastic leukemia alone or in combination with chemotherapy drugs pending further investigations on animal models as preclinical tests.
ABSTRACT
Embryonic cerebrospinal fluid [e-CSF] has an important role in development of embryonic and adult brain. Proteomic analysis suggests that this fluid has many morphogenes and cytokines that alter in time and space throughout embryonic life. The aim of this study was to evaluate the developmental effect of embryonic CSF on proliferation and differentiation of neuroprogenitor cells in different gestational age. In this experimental study, we examined the role of e-CSF on proliferation and differentiation of neuroprogenitor cells using neurosphere culture method. Neurospheres size analysis and MTT assay were used to assess cell proliferation after four days in vitro. Glial differentiation study was carried out by immunocytochemistry. Neurospheres size and percentage of glial fibrialy acidic protein [GFAP] positive cells were measured by image analyzer [image J]. The data were analyzed by one-way ANOVA, followed by the Tukey's post hoc test. Data were expressed as mean +/- SEM, and differences were considered significant when p<0.05, 0.01 and 0.001. Viability and proliferation of neuro progenitor cells in cultures conditioned with E16 CSF and E18 CSF were significantly increased compare to control group. A dramatic decrease in percentage of GFAP-positive cells was found following the application of CSF from E16 and E18 embryos, but not E20 CSF. Our data suggest that, e-CSF altered proliferation and differentiation of neuro progenitor cells in age dependent manner. E16 and E18 CSF enhanced proliferation and viability of neuro progenitor cells, and inhibited differentiation to glial fate in comparison with control group
Subject(s)
Animals, Laboratory , Neural Stem Cells , Cell Proliferation , Cell Differentiation , Embryonic Structures , Rats, Wistar , ImmunohistochemistryABSTRACT
Bee venom [BV], like many other complementary medicines, has been used for thousands of years for the treatment of a range of diseases. More recently, BV is also being considered as an effective composition for the treatment of cancer. Cancer is a major worldwide problem. It is obvious that the identification of compounds that can activate apoptosis could be effective on the treatment of cancer. BV is a very complicated mixture of active peptides, enzymes, and biologically active amines. The two main components of BV are melittin and phospholipase A[2] [PLA2]. Of these two components, melittin, the major active ingredient of BV, has been identified to induce apoptosis and to possess antitumor effects. We tried to review antineoplastic effects of BV in this study. The related articles were derived from different data bases such as PubMed, Elsevier Science, and Google Scholar using keywords including bee venom, cancer, and apoptosis. According to the results of this study, BV can induce apoptosis and inhibit tumor cell growth and metastasis. Results of in vivo experiments show that the anti-tumor effect of the BV is highly dependent on the manner of injection as well as the distance between the area of injection and the tumor cells. The results obtained from the reported studies revealed that BV has anticancer effects and can be used as an effective chemotherapeutic agent against tumors in the future
ABSTRACT
Complementary medicine uses bee venom [BV] to treat several diseases, including arthritis and skin diseases. BV contains mellitin, phospholipase A2, apamin and several other bioactive substances. According to the venom compounds, the purpose of this study was to examine the effects of BV on differentiation of K562cell line. In this experimental study, K562cells were treated with different doses of BV in different durations. BV toxicity was evaluated by MTT assay. Benzidine staining was used to investigate the effects of BV on K562 cell differentiation toward the erythroid line. In order to determine the type of cell death, annexin-V gene expression was analyzed by flow cytometry. Colony assay was used to measure BV ability in inhibiting colony formation. Morphological changes in the cells undergone treatment with BV were evaluated by wright-giemsa staining. MTT assay showed that bee venom with concentrations of 5.5-6 microg/ml and 3.5-4.5microg/ml result in 505 cell death in 24h and 48h, respectively. Morphological examination and benzidine staining showed that lower doses in longer period induce differentiation in these cells. Flow cytometry data showed significantly increased in annexin-V gene expression in cells which were treated with bee venom for 24 h. Colony assay demonstrated that the concentration of 1 microg/ml of BV results in 50% reduction in colony formation
ABSTRACT
Presumably, stem cells derived from mouse adipose tissue differentiate into lens fiber cells under induction by vitreous humor factors. Cataract is one of the most common ocular diseases that is highly treatable by replacing the ocular lens with artificial ones. The objective of this study was to find the natural lens replacements instead of the artificial ones. In this experimental study, stem cells from adipose tissue were obtained from inguinal fat pads from NMRI mouse. To demonstrate stemness potential of these cells, we used anti-OCT4, as stromal stemness markers with immunocytochemistry methods. This experiment took place for 14 days with different dosages of bovine vitreous humor. Express of ocular lens fiber cells markers [alpha crystalline] were detected by immunocytochemistry in experimental and control groups. All murine stem cells were positive to OCT4 marker which is used for stromal cells. Morphological studies showed that cells induced with 40% vitreous humor in culture media were locally longer and more aligned in parallel compared to control group cells. Also, the fiber like cells had large nuclei with multiple nucleoli that showed increase in production of proteins in those cells. The proof of the induction of these stem cells to lens fiber like-cells is the positive response of induced cells for crystalline markers in media with 40% vitreous humor by ANOVA test. Based on morphological appearance and positive response of experimental group to specific markers of lens fiber cells, it can be concluded that stem cells derived from mouse adipose tissue differentiate into lens fiber cells by treating them with vitreous humor
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There are several types of cancer, which cause millions of deaths worldwide every year. Many studies have confirmed that plants are adequate natural sources to be examined as anti-cancer drugs with fewer side effects than chemotherapy and radiotherapy. In this study the anti-cancer properties of Lavender aqueous extract on lymphocytes derived from patients with Hodgkin's lymphoma has been studied. In order to determine the cytotoxic effects of the extract on lymphocytes of patients in stages III and IV of Hodgkin's lymphoma and two different cell lines in the presence of different concentrations of aqueous extract of Lavender, MTT colorimetric assay and flow cytometry analysis were used. Findings indicated that Lavender inhibited cell proliferation in both lymphocytes and cell lines with different effects. The effective concentration of Lavender that decreased viability of Hodgkin's lymphoma cells below Lethal Concentration 50 [LC50] value was 100 micro g/ml and this was half of the therapeutic dose. In addition, apoptosis was the main mechanism the Hodgkin's lymphoma cell encountered when exposed to the aqueous extract of Lavender. Conclusion: This experiment proposes that aqueous Lavender extract can be regarded as a potential anti-cancer agent in future studies
Subject(s)
Humans , Plant Extracts , Hodgkin Disease , Lymphocytes , Cell Proliferation , Cell Line , CytotoxinsABSTRACT
Melatonin acts as an indirect antioxidant and is a powerful direct free radical scavenger and direct responses to melatonin in the gonads are detected. This study aims to investigate the influence of different doses of melatonin on preantral follicle development and oogenesis of in vitro cultured mouse ovarian follicles. Preantral follicles with diameters of 150- 175 micro m were mechanically isolated from NMRI mouse ovaries. Follicles were cultured in droplets of alpha-minimal essential medium [alpha-MEM] supplemented with 5% FBS, 100 mIU/ml rhFSH, 1%ITS, 100 IU/ml penicillin and 100 micro g/ml streptomycin in conjunction with varying doses of melatonin [0, 1, 10, 100 nM and 100, 500 pM] for six days. On day six, in vitro ovulation was induced by the addition of hCG/rEGF to the culture medium and after 16-20 h the maturation state of the oocytes was assessed. There was a significant [P<0.05] decrease in the number of surviving follicles in the groups that received 10, 100 nM and 500 pM melatonin compared to the other groups. After induction of in vitro ovulation, follicles in groups that received 1, 10, and 100 nM melatonin had higher ovulation rates [P<0.05] compared with the other groups. Oocyte maturation capacity was adversely influenced by five concentrations of melatonin and GV arrest was significantly higher compared to the control group [P<0.01]. Our data indicates that a dose of 100 pM melatonin has no toxic effects on follicular development and can be used to reduce oxidative stress in follicle culture systems
Subject(s)
Female , Animals, Laboratory , Ovarian Follicle/drug effects , In Vitro Oocyte Maturation Techniques , Oocytes/drug effects , Oogenesis/drug effects , Mice , OvulationABSTRACT
Multiple sclerosis [MS] is a progressive and autoimmune neurodegenerative disease of the central nervous system [CNS]. This disease is recognized through symptoms like inflammation, demyelination and the destruction of neurological actions. Experimental allergic encephalomyelitis [EAE] is a widely accepted animal model for MS. EAE is created in animals by injecting the tissue of myelin basic protein [MBP], CNS, or myelin oligodendrocyte glycoprotein [MOG] along with the adjuvant. EAE and MS are similar diseases. Honey Bee venom [Apis mellifera] contains a variety of low and high molecular weight peptides and proteins, including melittin, apamin, adolapin, mast cell degranulating peptide and phospholipase A2. Bee venom [BV] could exert anti-inflammatory and antinociceptive effects on the inflammatory reactions. The guinea pig spinal cord homogenate [GPSCH] is with the Complete Freund's Adjuvant [CFA], consisting of 1 mg/mL Mycobacterium tuberculosis. It was used for inducting EAE in Lewis rats for creating the MS model. The hematoxylin and eosin and luxol fast blue methods were used respectively in analyses of inflammation and detection of demyelination in the central nervous system. Furthermore, the ELISA and the high performance liquid chromatography [HPLC] were used for the assessment of tumor necrosis factor alpha [TNF-alpha] and nitrate in rats serum. In this study, we indicated that the treatment of EAE with Bee venom decreased the symptoms of clinical disorder, pathological changes, inflammatory cell infiltration, demyelination in the central nervous system, level of serum TNF-alpha, and the serum nitrates in rat EAE induced through GPSCH
ABSTRACT
Cyclooxygenase 2 is a key enzyme which converts arachidonic acid into prostaglandins. Cyclooxygenase 2 is triggered by inflammatory stimuli, such as cytokines. Its expression increases in tumors and Alzheimer's disease and ovarian hyperstimulation syndrome. Polycystic ovarian syndrome is a heterogeneous disease characterized by pathological angiogenesis and chronic anovulation. In the present study, the probable role of cyclooxygenase 2 in Wistar rats with polycystic ovarian syndrome was investigated. Thirty female Wistar rats [170-200 gr] were equally divided into three groups: 2 mg estradiol valerate was intramuscularly administered to each rat in the experiment group or group 1; the rats in group 2 were regarded as the sham group and received sesame oil injections and group 3 or the control group received no injections. After 60 days of treatment, animals were anaesthetized with chloroform and killed by decapitation. Ovaries were collected for histological and immunohistochemical evaluations. All the experiments were repeated three times. Morphologically, ovaries from the control group exhibited follicles in various stages of development and many fresh corpus luteum. In estradiol valerate group small follicles in early development were observed in addition to follicles showing evidence of atresia and many large cysts with thickened theca cell layer. Corpus luteum was rare or absent in group 2. The immunohistochemical analysis for cyclooxygenase 2 expression showed an increased expression of cyclooxygenase 2 enzyme in group 1. The results suggested the involvement of cyclooxygenase 2 in the progression to polycystic ovarian syndrome in a rat model
Subject(s)
Female , Animals, Laboratory , Polycystic Ovary Syndrome , Rats, Wistar , Estradiol/analogs & derivatives , Immunohistochemistry , Ovarian Follicle , Corpus LuteumABSTRACT
Background: Fetal cerebro- spinal fluid [CSF] contains many neurotrophic and growth factors. Rat pheochromocytoma PC12 cells have been widely used as an in vitro model of neuronal differentiation that undergo differentiation to sympathetic neuron-like cells in response to NGF, bFGF, EGF and GDNF
Materials and methods: CSF was removed by tapping the cisterna magna of Wistar rat fetuses [E17- E20]. PC12 cells were cultured in RPMI plus 10% FBS. The cell viability and cell proliferation were measured by MTT assay. Neural differentiation markers [MAP-2 and beta-III tubulin] expressions were analyzed by immunocytochemistry
Results: MAP-2 and beta-III tuobulin were expressed in PC12 cells cultured in CSF supplemented medium, but not in the cells from control cultures. Viability and cell proliferation were significantly elevated in PC12 cells cultured in CSF supplemented medium in E18 compared with control ones. A significant neuronal-like outgrowth appeared as early as Day 3 after the application of the CSF supplemented medium E17 and E19
Conclusion: Our data are in the same line with pervious studies that clarify crucial role of CSF neurotrophic factors in neuronal differentiation. Taken together we address PC12 neuronal differentiation to CSF induction by its components especially growth factors
ABSTRACT
Photodynamic Therapy [PDT] using 5-aminolevulinic acid [5-ALA]-induced protoporphyrin IX [PpIX] has been considered as a new method for treating neoplasms. However, ALA-PDT is suboptimal for thick tumors. Searching for new approaches, we investigated the effect of adding differentiation therapy [DT] with Vitamin E succinate [VES] to PDT in human prostate LN-CaP-FGC10 cancer cells in vitro. The purpose of DT was to reverse the lack of differentiation in cancer cells and to enhance the effectiveness of ALA- dependent PDT. Three groups of cells were grown on RPMI1640 culture medium supplemented with 10% FBS. The cells included: ALA-PDT cells, which received 0.3 mM ALA for 4 hours at dark, and exposed to 532 nm, 50 mW Nd-YAG laser beam for 3 min; DT+ALA-PDT cells, which received 6 micro g/ml VES for 24, 48 and 72 hours, followed by the addition of 0.3 mM ALA for 4 h and exposure to Nd-YAG laser beam for 3 min; control cells which were untreated. After 24 h, the percentage of cell viability was determined by MTT assay. Accumulation of PpIX was measured by spectrophotometry and fluorescent microscopy. Mechanism of induced cell death was investigated via Hoechst staining. The combination of both factors [VES and 5-ALA] lead to a significant increase in cell death after 72 h. Induction of differentiation augmented PpIX accumulation in cells treated with ALA. Elevated intracellular PpIX levels resulted in an enhanced lethal photodynamic sensitization of VES plus ALA-treated cells after 72 h. Apoptotic cell death by both ALA-PDT and VES-ALA-PDT was confirmed by Hoechst staining. Our data suggest that VES used in combination with 5-ALA may provide a new combinatorial approach for treating certain cancers