• Title/Summary/Keyword: Cancer cell line

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Identification of a Cancer Stem-like Population in the Lewis Lung Cancer Cell Line

  • Zhang, An-Mei;Fan, Ye;Yao, Quan;Ma, Hu;Lin, Sheng;Zhu, Cong-Hui;Wang, Xin-Xin;Liu, Jia;Zhu, Bo;Sun, Jian-Guo;Chen, Zheng-Tang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.3
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    • pp.761-766
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    • 2012
  • Objective: Although various human cancer stem cells (CSCs) have been defined, their applications are restricted to immunocompromised models. Developing a novel CSC model which could be used in immunocompetent or transgenic mice is essential for further understanding of the biomolecular characteristics of tumor stem cells. Therefore, in this study, we analyzed murine lung cancer cells for the presence of CSCs. Methods: Side population (SP) cells were isolated by fluorescence activated cell sorting, followed by serum-free medium (SFM) culture, using Lewis lung carcinoma cell (LLC) line. The self-renewal, differentiated progeny, chemosensitivity, and tumorigenic properties in SP and non-SP cells were investigated through in vitro culture and in vivo serial transplantation. Differential expression profiles of stem cell markers were examined by RT-PCR. Results: The SP cell fraction comprised 1.1% of the total LLC population. SP cells were available to grow in SFM, and had significantly enhanced capacity for cell proliferation and colony formation. They were also more resistant to cisplatin in comparison to non-SP cells, and displayed increased tumorigenic ability. Moreover, SP cells showed higher mRNA expression of Oct-4, ABCG2, and CD44. Conclusion: We identified SP cells from a murine lung carcinoma, which possess well-known characteristics of CSCs. Our study established a useful model that should allow investigation of the biological features and pharmacosensitivity of lung CSCs, both in vitro and in syngeneic immunocompetent or transgenic/knockout mice.

Effect of First Line Gastric Cancer Chemotherapy Regime on the AGS Cell Line - MTT Assay Results

  • Alizadeh-Navaei, Reza;Rafiei, Alireza;Abedian-Kenari, Saeid;Asgarian-Omran, Hossein;Valadan, Reza;Hedayatizadeh-Omran, Akbar
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.1
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    • pp.131-133
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    • 2016
  • Background: Combination chemotherapy regimes are common treatments for cancer. The aim of this study was to evaluation the effect of individual chemotherapeutic agents in comparison with a first line chemotherapy regime treatment in the AGS gastric cancer cell line by MTT assay. Materials and Methods: In this experimental study, AGS cells were grown in RPMI-1640 supplemented with 10% fetal calf serum and 100 IU/ml penicillin, and $10{\mu}g/ml$ streptomycinin, under a humidified condition at $37^{\circ}C$ with 5% CO2. All cells were washed with PBS and detached with trypsin, centrifuged and 8000 cells re-plated on to 96- well plates. LD50 doses of Epirubicin, Cisplatin and 5-fluorouracil were added to each well in mono or triple therapy. Anti-proliferative activities were determined by MTT assay after 24, 48 or 72 h. Results: Results of MTT assays showed that there were no significant differences among 3 drugs in monotherapy (p=0.088), but there was significant difference between combination therapy with epirubicin (P=0.031) and 5FU (p=0.013) on cell survival at 24 h. After 48 and 72 hours, cell viability showed significant differences between the 3 drugs (p=0.048 and P=0.000 for 48 and 72 h, respectively) and there was significant difference between combination therapy with epirubicin (P=0.035 and P=0.002 for 48 and 72 h, respectively). Conclusions: The results showed no significant differences between these chemotherapy drugs each given alone, but combination therapy with 3 drugs had significant effects on cell viability in comparison with epirubicin alone.

Oxidative Stress Induces Hypomethylation of LINE-1 and Hypermethylation of the RUNX3 Promoter in a Bladder Cancer Cell Line

  • Wongpaiboonwattana, Wikrom;Tosukhowong, Piyaratana;Dissayabutra, Thasinas;Mutirangura, Apiwat;Boonla, Chanchai
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3773-3778
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    • 2013
  • Increased oxidative stress and changes in DNA methylation are frequently detected in bladder cancer patients. We previously demonstrated a relationship between increased oxidative stress and hypomethylation of the transposable long-interspersed nuclear element-1 (LINE-1). Promoter hypermethylation of a tumor suppressor gene, runt-related transcription factor 3 (RUNX3), may also be associated with bladder cancer genesis. In this study, we investigated changes of DNA methylation in LINE-1 and RUNX3 promoter in a bladder cancer cell (UM-UC-3) under oxidative stress conditions, stimulated by challenge with $H_2O_2$ for 72 h. Cells were pretreated with an antioxidant, tocopheryl acetate for 1 h to attenuate oxidative stress. Methylation levels of LINE-1 and RUNX3 promoter were measured by combined bisulfite restriction analysis PCR and methylation-specific PCR, respectively. Levels of LINE-1 methylation were significantly decreased in $H_2O_2$-treated cells, and reestablished after pretreated with tocopheryl acetate. Methylation of RUNX3 promoter was significantly increased in cells exposed to $H_2O_2$. In tocopheryl acetate pretreated cells, it was markedly decreased. In conclusion, hypomethylation of LINE-1 and hypermethylation of RUNX3 promoter in bladder cancer cell line was experimentally induced by reactive oxygen species (ROS). The present findings support the hypothesis that oxidative stress promotes urothelial cell carcinogenesis through modulation of DNA methylation. Our data also imply that mechanistic pathways of ROS-induced alteration of DNA methylation in a repetitive DNA element and a gene promoter might differ.

Effect of Botulinum Toxin A on Proliferation and Apoptosis in the T47D Breast Cancer Cell Line

  • Bandala, Cindy;Perez-Santos, Jose Luis Martin;Lara-Padilla, Eleazar;Delgado Lopez, Ma. Guadalupe;Anaya-Ruiz, Maricruz
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.891-894
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    • 2013
  • The present study was performed to assess the activity of the botulinum toxin A on breast cancer cells. The T47D cell line was exposed to diverse concentrations of the botulinum toxin A and cell viability and apoptosis were estimated using MTT and propidium iodine/annexin V methods, respectively. Botulinum toxin A exerted greater cytotoxic activity in T47D cells in comparison with MCF10A normal cells; this appeared to be via apoptotic processes caspase-3 and -7. In conclusion, botulinum toxin A induces caspase-3 and -7 dependent apoptotic processes in the T47D breast cancer cell line.

Study on the Anti-cancer, Anti-metastasis and Immune Response Improvement of Aqua-acupuncture with Amomum Amarum Lourerio Infusion Solution (익지인약침(益智仁藥鍼)이 항암(抗癌) 및 면역기능(免疫機能)에 미치는 실험적(實驗的) 연구(硏究))

  • Park, Sang-Yong;Lee, Byung-Ryul
    • Journal of Acupuncture Research
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    • v.18 no.3
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    • pp.79-93
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    • 2001
  • Objective : This study was purposed to investigate the anti-cancer and anti-metastasis and immune response of Aqua-acupuncture with Amomum amarum Lourerio infusion solution. Methods : The Amomum amarum Lourerio infusion solution put into Chung-wan(CV12) of BALB/c or C57BL6 mice were rised to cancer by B15-F10 and HT1080, S-180 cancer cell line. Results : The following result have been obtained 1. The effect on expression of MMP-9 gene about the HT1080 cancer cell line was increased in 100, $50{\mu}g/m{\ell}$ diluent groups, compared with control group. 2. The effect on expression of MMP-9 gene about the B15-F10 cancer cell line was increased in all the sample groups, compared with control group. 3. S-180 cancer cell line transplants in BALB/c mice were inhibited significantly in weight inctease in all the sample groups, compared with control group. 4. The effect on spleen cell proliferation was decreased in all the sample groups, compared with control group. 5. The $IFN-{\gamma}$ in all the sample groups and the $TNF-{\alpha}$ in 25 and $12.5{\mu}g/m{\ell}$ diluent groups was increased. 6. In flow cytometry, the number of CD4+, CD19+ cell in all the sample group was increased and the number of CD8+ cell in $100{\mu}g/m{\ell}$ diluent group, compared with control group. Conclusion : According to the result, Aqua-acupuncture with Amomum amarum Lourerio infusion solution has significant anti-cancer, anti-metastasis and Immune response improvement.

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Talin-1 Correlates with Reduced Invasion and Migration in Human Hepatocellular Carcinoma Cells

  • Fang, Kun-Peng;Zhang, Jian-Lin;Ren, Yan-Hong;Qian, Ye-Ben
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.6
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    • pp.2655-2661
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    • 2014
  • Background: Talin-1 is a cytoskeleton protein that participates in cell migration and plays a role in tumor formation, migration, and metastasis in different types of cancer. Chinese investigators have observed that the levels of Talin-1 protein and mRNA expression in HCC tissues are significantly lower than in the adjacent non-cancerous tissue. However, Japanese investigators have reported that Talin-1 is upregulated in HCC. Tln2 as homologous gene of Tln-1, which encodes a very similar protein, but the role of Talin-2 is very little known in primary liver cancer (PLC). We investigated whether the expression of Talin-1 in PLC may be associated with the histological subtype as well as the role of Talin-1 in tumor cell invasion and migration using human hepatocellular carcinoma cell lines. Materials and Methods: We measured the mRNA expression levels of Talin-1 and Talin-2 in five human liver cancer cell lines and normal human liver cell ($LO_2$ cell line) by real-time PCR and the protein expression levels of Talin-1 by Western blot. Migration and invasion of the cells were assessed using transwell assays and cell scratch experiments, respectively, and proliferation was assessed by soft AGAR colony formation. Results: Talin-1 and Talin-2 expression differed significantly between the five human liver cancer cell lines and $LO_2$ cell line (p<0.05). Compared with the $LO_2$ cell line, the invasion and migration capabilities of the five cancer cell lines differed significantly (p<0.05). Similarly, the colony-forming ability differed (p<0.05). Conclusions: High levels of Talin-1 expression are correlated with reduced invasion and migration as well as decreased malignancy in human liver cancer cell lines; the suppression of Talin-1 promotes invasion and migration. In addition, Talin-2 may be correlated with invasion and migration in human hepatocellular carcinoma.

Synaptic Vesicle Protein 2 (SV2) Isoforms

  • Bandala, Cindy;Miliar-Garcia, A.;Mejia-Barradas, C.M.;Anaya-Ruiz, M.;Luna-Arias, J.P.;Bazan-Mendez, C.I.;Gomez-Lopez, M.;Juarez-Mendez, S.;Lara-Padilla, E.
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.5063-5067
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    • 2012
  • New molecular markers of cancer had emerged with novel applications in cancer prevention and therapeutics, including for breast cancer of unknown causes, which has a high impact on the health of women worldwide. The purpose of this research was to detemine protein and mRNA expression of synaptic vesicle 2 (SV2) isoforms A, B and C in breast cancer cell lines. Cultured cell lines MDA-MB-231, SKBR3, T47D were lysed and their protein and mRNA expression analyzed by real-time PCR and western blot technique, respectively. SV2A, B proteins were identified in non-tumor (MCF-10A) and tumor cell lines (MDA-MB-231 and T47D) while SV2C only was found in the T47D cell line. Furthermore, the genomic expression was consistent with protein expression for a such cell line, but in MDA-MB-231 there was no SV2B genomic expression, and the SV2C mRNA and protein were not found in the non tumoral cell line. These findings suggest a possible cellular transdifferentiation to neural character in breast cancer, of possible relevance to cancer development, and point to possible use of SV2 as molecular marker and a vehicle for cancer treatment with botulinum toxin.

Cell Growth Inhibitory Effect of Tissue Cultured Root of Wild Panax ginseng C.A. Mayer Extract on Various Cancer Cell Lines

  • Park, Jeong-Sook;Lee, Tae-Woong;Han, Kun
    • Natural Product Sciences
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    • v.15 no.1
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    • pp.1-7
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    • 2009
  • This study was performed to investigate the cell growth inhibitory effect of tissue cultured root of wild Panax ginseng C.A. Mayer (tcwPG). The human stomach carcinoma cell line, MKN 74, was incubated with 70% EtOH extract of tcwPG or Panax ginseng C.A. Mayer (PG) for 24 hrs. tcwPG inhibited cell growth at a concentration of $250{\mu}g/ml$. However, Panax ginseng extract did not inhibit cell growth at the same concentration. We also tested the ethyl acetate and $H_2O$ fractions of tcwPG. The inhibitory effect of the ethyl acetate fraction on cell proliferation in MKN 74 cells was more potent than that of the crude extract, and the inhibitory effect of the $H_2O$ fraction was less than that of the ethyl acetate fraction. When we separated tcwPG into polar and non-polar saponin fractions and then measured cell growth inhibition, the non-polar saponin in tcwPG exhibited cytotoxicity. To compare the effects of tcwPG on various cancer cell lines, we measured cytotoxicity in MKN 74 (stomach cancer cell line), SW 620 (colon cancer cell line) and PC 3 (prostate cancer cell line). All three cell lines showed cell growth inhibition, and the cell growth inhibitory effects were not quite different in the various cell lines. The non-polar saponins of tcwPG arrested PC 3 cells at G1-phase as did Panax ginseng.

IN-VITRO STUDY OF CO2 EXTRACT OF TERMINALIA CHEBULA IN BREAST CANCER CELL LINE MD-MBA-231

  • Chandil, Shachi;Bamoriya, Harikishan;More, D.B.
    • CELLMED
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    • v.11 no.3
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    • pp.16.1-16.7
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    • 2021
  • Cancer is an abnormal growth of cells in body which leads to death. These cells are born due to imbalance in cell proliferation mechanism. In 2018, WHO released new statistics on cancer incidence, mortality, and prevalence worldwide i.e., GLOBOCAN 2018 estimates for 28 types of cancer in which more prevalence of cervix and breast cancer. According to survey, in India about 7.8 million cancer deaths and 11.5 million new cases arise in 2018, which will increase to 19.3 million new cases per year by 2025. Though breast cancer as such is not explained anywhere in Ayurvedic compendia, correlations can be done with the Stana Arbuda. Ayurveda, the ancient system of medicine came into existence 1000's of years ago with an objective of maintaining the health of people and treating diseases. Many herbs used in Ayurveda have been screened for activity against cancer and in-vitro and in-vivo studies have given promising leads. The plant, called as "Mother of Medicine", Haritaki has been extensively studied for its various ailments because of its extraordinary healing potency. Haritaki (Terminalia chebula Retz.), Family: Combretaceae have a great therapeutic value and is widely distributed in India. Dried fruit of Terminalia chebula contains high quantities phenolic compounds consist of ellagic acid, gallic acid and chebulic acid. The fruit extract of T. chebula is having different biological properties like anticancer, antioxidant, hepatic and renal protective activities etc. In this study, we focus on the use of CO2 extract of Terminalia chebula, on the breast cancer cell line MDA-MB-231. All tests proved that CO2 extract of Terminalia chebula containing active chemical component, therefore our experiment showed the positive results for CO2 extract of Terminalia chebula against breast cancer cell line cancer MDA-MB-231. The MTT assay results were used to evaluate the anti-cancer activity of the extract. The percentage of cell growth and cell viability were calculated from tabulated result values of MTT assay. Cell viability MTT assay also showed significant growth inhibition, at the same time statistical analysis of MTT assay also proved significant results.

Combined Effects of Curcumin and Triptolide on an Ovarian Cancer Cell Line

  • Cai, Ying-Ying;Lin, Wei-Ping;Li, Ai-Ping;Xu, Jian-Yang
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4267-4271
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    • 2013
  • Background: As natural medicines in Asia, curcumin and triptolide extracted from different drug plants have proven to possess anticancer potential and widely used for anti-cancer research. The present study attempted to clarify that curcumin and triptolide synergistically suppress ovarian cancer cell growth in vitro. Methods: To test synergic effects, cell viability and apoptosis were analyzed after curcumin and triptolide combination treatment on ovarian cancer cell lines. Synergistic effects on apoptosis induction were determined by lactate dehydrogenase (LDH) leakage assay, intracellular reactive oxygen species (ROS) assay, mitochondrial membrane potential (MMP) loss assay and flow cytometry analysis. Critical regulators of cell proliferation and apoptosis related were analyzed by qRT-PCR and Western blotting. Results: We showed that the combination of curcumin and triptolide could synergistically inhibit ovarian cancer cell growth, and induce apoptosis, which is accompanied by HSP27 and HSP70, indicating that HSP27 and HSP70 play the important role in the synergic effect. Conclusions: From the result present here, curcumin and triptolide combination with lower concentration have a synergistic anti-tumor effect on ovarian cancer and which will have a good potential in clinical applications.