scholarly journals Cytotoxicity, Apoptosis induction and change of p53, PARP, p21 and Bcl-2 genes expression in the human anaplastic thyroid carcinoma cells line (SW-1736) with curcumin

2021 ◽  
Vol 5 (1) ◽  
pp. 10
Author(s):  
Khalil Khashei ◽  
Parinaz Tavakoli ◽  
Leila Rouhi ◽  
Somayee Raisi
Keyword(s):  
Thyroid Carcinoma ◽  
Cell Line ◽  
Annexin V ◽  
Poor Response ◽  
Anaplastic Thyroid Carcinoma ◽  
Control Group ◽  
Dependent Manner ◽  
Apoptosis Induction ◽  
Thyroid Cells ◽  
Genes Expression

Anaplastic thyroid carcinoma is highly invasive with a poor response to a treatment. In this study, curcumin bioavailability and its effects on apoptosis induction and selected genes expression of the human anaplastic thyroid carcinoma cell line (SW-1736) were examined. SW-1736 cells were incubated for 24 and 48 hours with different concentrations of curcumin 2.5, 5, 7.5 and 10 μM to examine bioavailability, and for 24, 48 and 72 hours with concentrations of 2.5, 5, 7.5, 10 μM and 2.5, 5 and 10 μM respectively to examine apoptosis and the expression of p53, PARP, p21 and Bcl-2 genes. Then, bioavailability was analyzed by MTS kit, apoptosis was analyzed by flow- cytometry using Annexin V-FITC/PI kit and the expression of p53, PARP, p21 and Bcl-2 genes were analyzed by Real Time PCR. ANOVA test and SPSS 16 software were used for statistical analysis. The results indicate that curcumin at the concentration of 7.5 μM has significantly decreased bioavailability in anaplastic thyroid cells in comparison with other treatments at both incubation periods. Induction of apoptosis with increasing concentration of curcumin in dose and time dependent manner increased in this cell line. Also, treatment with curcumin significantly decreased the expression of Bcl-2 gene and increased the expression of p53, PARP and p21 genes in some experimental groups compared to the control group. Curcumin inhibited the growth, proliferation and invasion of anaplastic thyroid cancer cells through altering the expression of the genes involved in the apoptosis process

Inhibition Effects of baicalin on Lymphoma Cell Line CA46 in Vitro and in Vivo

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 5053-5053
Author(s):  
Jian Da Hu ◽  
Yi Huang ◽  
Yingyu Chen ◽  
Tiannan Wei ◽  
Tingbo Liu ◽  
...  
Keyword(s):  
Cell Line ◽  
Biological Effects ◽  
Annexin V ◽  
Lymphoma Cell ◽  
Control Group ◽  
Dependent Manner ◽  
Lymphoma Cell Line ◽  
Proliferation Inhibition

Abstract Baicalin is a traditional Chinese medicine with multiple biological effects. Some researches showed baicalin has anti-tumor effects in solid tumor, such as prostate cancer. In order to investigate its effects on proliferation inhibition and apoptosis induction in human lymphoma cell, we treated Burkitt lymphoma cell line CA46 with baicalin in vitro and in vivo of CA46 xenograft. Baicalin remarkably inhibited the cell proliferation, with an IC50 value of 10μM. Apoptosis was remarkably induced by baicalin in a dose-dependent manner, which was detected by Annexin V FITC/PI double staining analysis, TUNEL labeling method and DNA fragmentation respectively. Furthermore, RT-PCR showed that the mRNA expressions of c-myc and bcl-2 in treated CA46 cell decreased in a time-dependent manner. Western-Blot showed that the protein expressions of c-myc, bcl-2, procaspase-3 and PARP(116KD) in baicalin treated CA46 cell were down-regulated, while the expression of PARP(85KD) increased. Based on the results in vitro, we investigated in vivo efficacy of baicalin, alone or in combination with cytotoxic drug VP16, for treatment in CA46 nude mice xenograft. Baicalin with the dosage of 40mg/kg/d and 80kg/mg/d could remarkably inhibit the growth of the tumor compared with control group. Combination of baicalin and VP16 had better anti-tumor effects. Histological examination of tumor samples showed more necrotic cells in treated groups. And obvious apoptosis could be observed by electron microscope. No adverse events were found in treated groups. From above we could conclude that baicalin could efficiently induce proliferation inhibition and apoptosis of CA46 cells in vitro and in vivo, which may be related with the down-regulation of c-myc and bcl-2 expressions, as well as the up-regulation of caspase-3 activity.

scholarly journals Clodronate alleviates cachexia and prolongs survival in nude mice xenografted with an anaplastic thyroid carcinoma cell line

2006 ◽  
Vol 190 (2) ◽  
pp. 415-423 ◽  
Author(s):  
Cheng-Hsu Wang ◽  
Yung-Chi Shen ◽  
Jia-Juan Hsieh ◽  
Kun-Yun Yeh ◽  
John Wen-Cheng Chang
Keyword(s):  
Thyroid Carcinoma ◽  
Cell Line ◽  
Nude Mice ◽  
Cancer Cachexia ◽  
Carcinoma Cell ◽  
Anaplastic Thyroid Carcinoma ◽  
Carcinoma Cell Line ◽  
Dependent Manner ◽  
Thyroid Carcinoma Cell Line ◽  
Thyroid Carcinoma Cell

Cancer cachexia is one of the most common manifestations of advanced malignant disease and is frequently associated with decreased survival. Previously, we reported the establishment of a new anaplastic thyroid carcinoma cell line, Thena, and its mouse xenograft, Thena-Nu, which induced cachexia in athymic nude mice. Subsequent studies showed that the addition of clodronate to Thena-Nu cultures reduced cell proliferation as well as cytokine production in a dose- and time-dependent manner. Weekly administration of clodronate induced tumor cytostasis, attenuation of cachexia, as well as prolongation of survival in Thena-Nu-bearing mice. Reduced serum interleukin 6, tumor necrosis factor-α, and granulocyte colony stimulating factor levels were detected, whereas, serum leukemia inhibitory factor levels were not reduced. Liver necrosis, observed in tumor-bearing mice, was also improved following clodronate treatment. Discontinuation of clodronate treatment, however, resulted in progressive tumor growth and weight loss. Our results demonstrated that clodronate could exert therapeutic efficacy on amelioration of cancer cachexia in the hosts. Nevertheless, this study also points out that a longer period of treatment is required to maintain these effects.

Ruxolitinib Regulates the Autophagy Machinery in Multiple Myeloma Cells

2020 ◽  
Vol 20 (18) ◽  
pp. 2316-2323 ◽  
Author(s):  
Alican Kusoglu ◽  
Bakiye G. Bagca ◽  
Neslihan P.O. Ay ◽  
Guray Saydam ◽  
Cigir B. Avci
Keyword(s):  
Multiple Myeloma ◽  
Cell Line ◽  
B Lymphocyte ◽  
Plasma Cells ◽  
Annexin V ◽  
Myeloma Cell ◽  
Control Group ◽  
Multiple Myeloma Cell ◽  
Ic50 Values ◽  
Stat Pathway

Background: Ruxolitinib is a selective JAK1/2 inhibitor approved by the FDA for myelofibrosis in 2014 and nowadays, comprehensive investigations on the potential of the agent as a targeted therapy for haematological malignancies are on the rise. In multiple myeloma which is a cancer of plasma cells, the Interleukin- 6/JAK/STAT pathway is emerging as a therapeutic target since the overactivation of the pathway is associated with poor prognosis. Objective: In this study, our purpose was to discover the potential anticancer effects of ruxolitinib in ARH-77 multiple myeloma cell line compared to NCI-BL 2171 human healthy B lymphocyte cell line. Methods: Cytotoxic effects of ruxolitinib in ARH-77 and NCI-BL 2171 cells were determined via WST-1 assay. The autophagy mechanism induced by ruxolitinib measured by detecting autophagosome formation was investigated. Apoptotic effects of ruxolitinib were analyzed with Annexin V-FITC Detection Kit and flow cytometry. We performed RT-qPCR to demonstrate the expression changes of the genes in the IL-6/JAK/STAT pathway in ARH-77 and NCI-BL 2171 cells treated with ruxolitinib. Results: We identified the IC50 values of ruxolitinib for ARH-77 and NCI-BL 2171 as 20.03 and 33.9μM at the 72nd hour, respectively. We showed that ruxolitinib induced autophagosome accumulation by 3.45 and 1.70 folds in ARH-77 and NCI-BL 2171 cells compared to the control group, respectively. Treatment with ruxolitinib decreased the expressions of IL-6, IL-18, JAK2, TYK2, and AKT genes, which play significant roles in MM pathogenesis. Conclusion: All in all, ruxolitinib is a promising agent for the regulation of the IL-6/JAK/STAT pathway and interferes with the autophagy mechanism in MM.

Antiproliferative and Apoptotic Properties of Andrographolide Against Human Colon Cancer DLD1 Cell Line

2020 ◽  
Vol 20 (6) ◽  
pp. 930-942 ◽  
Author(s):  
Imran Khan ◽  
Sadaf Mahfooz ◽  
Irfan A. Ansari
Keyword(s):  
Colon Cancer ◽  
Cell Line ◽  
Mtt Assay ◽  
Caspase 3 ◽  
Chemotherapeutic Agents ◽  
Synergistic Activity ◽  
Dependent Manner ◽  
Apoptosis Induction ◽  
Dld1 Cell ◽  
Activation Assay

Background: In recent years, natural products have received great attention for cancer prevention owing to their various health benefits, noticeable lack of toxicity and side effects, and the limitations of chemotherapeutic agents. Andrographolide, a labdane diterpenoid is a principal bioactive constituent of Andrographis paniculata Nees, exhibits significant anticancer activity. Objective: The efficacy of andrographolide on colon cancer cells is yet to be elucidated completely. Therefore, we investigated the anticancer efficiency of andrographolide in colon cancer DLD1 cell line. Methods: Antiproliferative activity of andrographolide on DLD1 cells was evaluated by MTT assay, LDH release assay, morphological analysis and colony formation assay. Induction of apoptosis was determined by DAPI staining, Annexin V-FITC staining assay, and caspase-3 activation assay. Role of andrographolide induced cellular reactive oxygen species (ROS) and its association with apoptosis induction in DLD1 cells was elucidated by DCFDA dye. Synergistic ability of andrographolide with 5- fluorouracil (5-FU) and paclitaxel (PTX) was evaluated by MTT assay. Results: Results of the present study indicated that andrographolide declined cell viability of DLD1 cells in a concentration and time-dependent manner. Andrographolide induced apoptosis via nuclear condensation, phosphatidylserine externalization and caspase-3 activation. It also augmented cellular ROS levels which were in turn associated with apoptosis induction in DLD1 cells. Moreover, andrographolide displayed synergistic activity with 5-FU and PTX against DLD1 cells. Conclusion: The present study showed that andrographolide demonstrated antiproliferative and apoptotic properties, moreover it also displayed synergistic effect with chemotherapeutic drugs in colon cancer DLD1 cells.

scholarly journals Cytotoxic Effects of Pistacia Atlantica (Baneh) Fruit Extract on Human KB Cancer Cell Line

2019 ◽  
Vol 62 (1) ◽  
pp. 30-34
Author(s):  
Zohreh Jaafari-Ashkvandi ◽  
Saeed Yousefi Shirazi ◽  
Somayeh Rezaeifard ◽  
Azadeh Hamedi ◽  
Nasrollah Erfani
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Normal Fibroblast ◽  
Human Gingival Fibroblast ◽  
Gingival Fibroblast ◽  
Dependent Manner ◽  
Apoptosis Induction ◽  
Cytotoxic Effects ◽  
Induction Of Apoptosis

Plants with anticancer properties are considered as cancer preventive and treatment sources, due to their some biological effects. Apoptosis induction and anti-proliferative effects of Baneh extract on various cancer cell lines have been reported. Hence, this study was designed to evaluate the cytotoxic effects of this fruit on KB and human gingival fibroblast cell lines (HGF). KB and HGF cells were treated with various concentrations of ethanolic Baneh extract and cisplatin as positive control. Cytotoxic activity and apoptosis induction were investigated using WST-1 and Annexin V assays. Data were analyzed using ANOVA and student’s t-tests. IC50 after 24 and 48 hours treatment were respectively 2.6 and 1 mg/mL for KB cell line, and 1.5 and 1.6 mg/mL for HGF cell. During 48 hours Baneh extract induced apoptosis without significant necrosis, in a time- and dose-dependent manner. The induction of apoptosis in KB cells was significantly higher than HGF. It seems that ethanolic extract of Baneh contains compounds that can suppress KB cell growth through the induction of apoptosis. Within 48 hours, less cytotoxic effects were observed on normal fibroblast cells; therefore, it might be a potential anticancer agent.

scholarly journals Emerging Effects of Sepantronium Bromide (YM155) on MOLT-4 Cell Line Apoptosis Induction and Expression of Critical Genes Involved in Apoptotic Pathways

2019 ◽  
Vol 10 (1) ◽  
pp. 81-87
Author(s):  
Kobra Shojaei Moghadam ◽  
Majid Farshdousti Hagh ◽  
Mohammad Reza Alivand ◽  
Masoumeh Fardi ◽  
Ali Akbar Movassaghpour ◽  
...  
Keyword(s):  
Cell Line ◽  
Epithelial To Mesenchymal Transition ◽  
Target Genes ◽  
Lymphoblastic Leukemia ◽  
Annexin V ◽  
Apoptosis Induction ◽  
Mesenchymal Transition ◽  
Expression Levels ◽  
Apoptotic Pathways ◽  
Sepantronium Bromide

Purpose: Sepantronium bromide (YM155) is a Survivin inhibitor which recently advanced as an anticancer agent in phase II clinical trials. Survivin belongs to IAP (inhibitor of apoptosis) gene family and is a pivotal target for treatment due to its overexpression and oncogenic function in many malignancies, including acute lymphoblastic leukemia (ALL). Although survivin is a specific target for YM155, recent reports have shown that it has many other crucial targets that regulate its anti-apoptotic effects. The aim of this study was to investigate whether YM155 could have an effect on cell death-inducing genes as well as inducing apoptosis in T-ALL MOLT4- cell line. Methods: We treated MOLT-4 cells with increasing concentrations of YM155 and then cell viability was determined using MTT (methyl thiazolyl tetrazolium) assay. Also, the rate of induction of apoptosis in MOLT-4 cells and the target genes expression levels were evaluated by Annexin V/PI and real-time PCR, respectively. Results: YM155 inhibited cell growth in MOLT-4 cells. This outcome is achieved by inducing apoptosis and a significant increase in the expression level of P53, MiR-9, caspase 3 and decreasing the mRNA expression levels of survivin, Sirtuin1(SIRT1), member of anti-apoptotic proteins family (Bcl-2), and epithelial-to-mesenchymal transition (EMT) initiating factors Snail1and Zeb2. Conclusion: The results showed that use of YM155 can be a potential drug therapy in T-ALL patients with promising effects on apoptosis induction.

Anti-Leukemic Property of Sulphoraphane In Precursor B Cell Acute Lymphoblastic Leukemia

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3974-3974
Author(s):  
Koramit Suppipat ◽  
Xiao Zhu ◽  
Chun Shik Park ◽  
H. Daniel Lacorazza
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
B Cells ◽  
B Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Lymphoblastic Leukemia ◽  
Annexin V ◽  
Dependent Manner ◽  
Carcinogenic Activity ◽  
Childhood All

Abstract Abstract 3974 Acute lymphoblastic leukemia (ALL) is the most common form of hematologic malignancy in children. In spite of significant advances achieved in the treatment of childhood ALL, one fifth of these patients still relapse after the standard treatment. Moreover, relapse ALL is the second most common cause of cancer-related deaths in children. The development of novel therapies is prevented by a limited understanding of the exact pathobiology. There are emerging evidences that the transcription factor KLF4 has a tumor suppressor property in ALL. Recently, a gene expression classifier study in pediatric precursor B-cell ALL (pre-B ALL) showed that KLF4 expression was significantly reduced in high risk ALL patients with positive MRD after induction. This finding suggests a possible role of this cell cycle inhibitor on the development, expansion and drug-resistant of leukemic cells. Several studies demonstrate that overexpression of KLF4 in normal B cells and BCR transformed B cells show increased apoptosis and reduced proliferation. Furthermore, we recently described that KLF4 inhibits proliferation of naïve lymphocytes by activating p21 (Yamada, et al, 2009). Sulphoraphane (SF; 4-methylsulfonylbutyl isothiocyanate) is a dietary compound derived from Cruciferae vegetables with anti-carcinogenic activity in colon cancer by upregulating KLF4 and p21 among other genes. Thus, we hypothesized that SF could also exhibit anti-leukemic activity in human-derived acute lymphoblastic leukemia cells via the activation of KLF4. The pre-B ALL cell lines (Nalm6, REH, RS-4, SUP-B15) and an EBV transformed B cell line were treated with different concentrations of SF (0-40 μM) for 24–48 hours. Then, cell number was estimated using an ATP-based viability method. Flow cytometric analysis of ANNEXIN-V/7-AAD binding as well as CFSE dilution was used to measure apoptosis and proliferation respectively. We found that SF induced cytotoxicity in Nalm-6, REH and RS-4 cell lines in a dose and time dependent manner. This cytotoxic effect was less pronounced in EBV-transformed B cell line. SF treatment led to increased ANNEXIN-V and 7-AAD positive cells (82% apoptotic cells in SF-treated versus 9% in DMSO control). Further, SF-treated cells displayed significantly less proliferation in comparison to DMSO controls thus suggesting that SF inhibits cellular proliferation. Preliminary data also suggest that SF-mediated apoptosis is caused by upregulation of KLF4. In conclusion, Sulphoraphane exhibits an anti-leukemic property by inducing apoptosis and abrogating proliferation in pre-B ALL cell lines. Thus, sulphoraphane could potentially be used as an adjunctive therapy in a subgroup of pre-B ALL patients who have decreased expression of KLF4. Disclosures: No relevant conflicts of interest to declare.

Study on the Effects of Rapamycin Reverse AMN107 resistance on k562/AO2 Cells

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 5017-5017
Author(s):  
Jishi Wang ◽  
Chang Yang ◽  
Cheng Chen ◽  
Qin Fang
Keyword(s):  
Western Blot ◽  
Cell Line ◽  
Chronic Myelogenous Leukemia ◽  
Copy Number ◽  
Caspase 3 ◽  
Annexin V ◽  
Myelogenous Leukemia ◽  
Control Group ◽  
Rt Pcr ◽  
Regulation Of Expression

Abstract Abstract 5017 Aim Rapamycin(RAPA) is the inhibitor of m-TOR. The activation of m-TOR results in changes in multiple cellular processes, eg, catabolism, anabolism, proliferation, growth and apoptosis. Rapamycin, which directly inhibits the Akt downstream molecule mammalian target of rapamycin (mTOR), effectively inhibits survival and proliferation in the development of BCR- ABL–induced chronic myelogenous leukemia(CML) from PTENfl/fl. Nilotinib (AMN107), a BCR-ABL tyrosine kinase inhibitor, was developed to surmount resistance or intolerance to imatinib in patients with Philadelphia positive chronic myelogenous leukemia. We aim to introduce the CML cell line (k562/A02) resistant to AMN107 and investigate the effect of RAPA reverse AMN107 resistance on k562/A02 cell line. Method K562/A02 be added in the culture medium final concentration 1μmol/ L of AMN107 drugs a week before the withdrawal. For all experiments, cells were on the logarithmic phase.K562/A02 cells were treated with 400nmol/L rapamycin(400nmol/L RAPA group),100nmol/L rapamycin(100nmol/L RAPA group), 20μmol/L nilotinib(20μmol/L AMN107 group), 400nmol/L rapamycin +20μmol/L nilotinib (400nmol/L RAPA+20μmol/L AMN107 group), 100nmol/L rapamycin +20μmol/L nilotinib (100nmol/L RAPA+20μmol/L AMN107 group). Tetrazolium—based colorimetric assay (MTT) was used to detect the inhibiton effect of cell growth. The expression of Survivin, m-TOR, Bcl-2, and Caspase-3 were detected by western blot; Cell apoptosis was inspected by Annexin V/PI double staining method; The expression of Survivin, m-TOR, Bcl-2 and Caspase-3 in mRNA level was determined using reverse transcriptase—polymerase chain reaction (RT-PCR). BCR-ABL gene was detected by Real Time-PCR when K562/AO2 cells in different groups were treated by drugs for 48h. Result K562/A02 cells were successfully introduced to resistant toAMN107. Combination of 400 and 100 nmol/L RAPA with AMN107 could significantly enhance the sensitivity of k562/A02 to AMN107. The reverse factor was 1.97 and 2.73 fold respectively. RT-PCR and Western blot method indicated that the expression of Survivin and Bcl-2 was increased by RAPA+AMN107 group, but the MDR1 and Caspase-3 was decreased. The expression of BCR-ABL fusion gene in100 nmol/L RAPA group was higher than the blank groups, the copy number is 5.6×10−2. But the copy number in 400nmol/L RAPA +20μmol/L AMN107 group is1.64×10−2 that is lowest. Annexin V/PI showed that the apoptosis rates were (8.21±0.05)%, (32.07±0.03)%, (28.88±0.11)%, (16.57±0.05)%, (38.49±0.04)%, and(42.31±0.07)% respectively in control group, AMN107 group, 400nmol/L RAPA group, 100nmol/L RAPA group, 100nmol/L RAPA +20μmol/L AMN107 group, and 400nmol/L RAPA +20μmol/L AMN107 group. Conclusion We successfully introduced K562/A02 resistant toAMN107. Moreover, RAPA can enhance the sensitivity of AMN107 resistant 562/A02 cells. Combined of RAPA and AMN107 has significant synergistic growth inhibiting effect and apoptosis inducing effect on AMN107-resistant K562/A02 cells. The mechanism may be related at down-regulation of expression MDR1 and up-regulation of expression Survivin. Disclosures: No relevant conflicts of interest to declare.

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