• Title/Summary/Keyword: Hela

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Study of Apoptosis by Scirpi Tuber in Hela Cell and MCF-7 Cell (자궁경부암(子宮頸部癌)과 유방암(乳房癌)에 대한 삼릉(三稜)의 세포자멸사 연구)

  • Yoo, Gap-Soon;Lee, Jin-Moo;Lee, Chang-Hoon;Jang, Jun-Bock;Lee, Kyung-Sub
    • The Journal of Korean Obstetrics and Gynecology
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    • v.24 no.3
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    • pp.1-13
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    • 2011
  • Objectives: This study was designed to investigate the analysis of apoptosis by Scirpi Tuber in Hela cell and MCF-7 cell. Methods: For cytotoxic effect of Scirpi Tuber extract, Scirpi Tuber extract were cultured on NIH3T3 cell in vitro. After treatment with various concentration of Scirpi Tuber, cell growth was evaluated in Hela cell and MCF-7 cell. Hoechst 33342 staining was performed to estimate DNA fragment effect of Scirpi Tuber on the apoptosis in Hela cell and MCF-7 cell. Annexin V/PI apoptosis assay was used to estimate the effects of Scirpi Tuber on the early apoptosis in Hela cell MCF-7 cell. All the stained cells were analyzed by a FACS. RT-PCR was used to estimate the apoptosis gene expression effect of Scirpi Tuber extract on Hela cell and MCF-7 cell. Results: Cytotoxic effect of Scirpi Tuber extract was not found on per NIH3T3 cell. The viability of Hela cell was significantly decreased Scirpi Tuber (500, $1000{\mu}g/m\ell$) in Hela cell 1day, 3day and 5days after treatment (p<0.01). The viability of MCF-7 cell was significantly decresed Scirpi Tuber ($1000{\mu}g/m\ell$) in MCF-7 cell (p<0.01), Scirpi Tuber ($500{\mu}g/m\ell$) in MCF-7 cell only 3days after treatment (p<0.01). In RT-PCR analysis, after treatment of $100{\mu}g/m\ell$ of ACR extract, BCL-2 were decreased and BAX, caspase-3 were increased both in Hela cell and MCF-7 cell. DNA fragmentation was observed the Scirpi Tuber on Hela cell and MCF-7 cell. As time goes on DNA fragmentation incresed. In Annexin V/PI apoptosis assay, after treatment of $1mg/m\ell$ of Scirpi Tuber, the early apoptotic cell increased both in Hela cell and MCF-7 cell. As time goes on apoptotic cell increased. Conclusion: Scirpi Tuber appears to have considerable activity on the apoptosis in Hela cell and MCF-7 cell.

RASSF1A Suppresses Proliferation of Cervical Cancer Cells

  • Feng, Lei;Li, Jie;Yan, Ling-Di;Tang, Jian
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.14
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    • pp.5917-5920
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    • 2014
  • Background: This study aimed to explore the effects of ras association domain family 1 A (RASSF1A) on proliferation and apoptosis of human cervical cancer cell line Hela cells. Materials and Methods: RASSF1A was cloned into the pcDNA3.1(+) vector to generate pcDNA3.1(+)-RASSF1A plasmid for transfection into Hela cells. Changes in the proliferation and apoptosis of cultured Hela cells were examined by the 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium chloride assay and flow cytometry. A protein array was used to analyze the expression of apoptotic factors. Results: Plasmid pcDNA3.1(+)-RASSF1A was generated and transfected into Hela cells to stably express RASSF1A in Hela cells. RASSF1A transfection was effective in inhibiting the proliferation of Hela cells up to 52.4%, as compared to cells transfected with an empty plasmid. RASSF1A expression also successfully induced apoptosis in human cervical cells with an apoptosis rate of 20.5%. More importantly, protein array results showed that RASSF1 A transfection induced overexpression of p21 and caspase 8, while decreasing the expression of survivin in Hela cells. Conclusions: RASSF1A expression was effective in suppressing the proliferation and increasing apoptosis of Hela cells, and may be a potential therapy for cervical cancer in clinic.

Solution structure and functional analysis of HelaTx1: the first toxin member of the κ-KTx5 subfamily

  • Park, Bong Gyu;Peigneur, Steve;Esaki, Nao;Yamaguchi, Yoko;Ryu, Jae Ha;Tytgat, Jan;Kim, Jae Il;Sato, Kazuki
    • BMB Reports
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    • v.53 no.5
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    • pp.260-265
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    • 2020
  • Scorpion venom comprises a cocktail of toxins that have proven to be useful molecular tools for studying the pharmacological properties of membrane ion channels. HelaTx1, a short peptide neurotoxin isolated recently from the venom of the scorpion Heterometrus laoticus, is a 25 amino acid peptide with two disulfide bonds that shares low sequence homology with other scorpion toxins. HelaTx1 effectively decreases the amplitude of the K+ currents of voltage-gated Kv1.1 and Kv1.6 channels expressed in Xenopus oocytes, and was identified as the first toxin member of the κ-KTx5 subfamily, based on a sequence comparison and phylogenetic analysis. In the present study, we report the NMR solution structure of HelaTx1, and the major interaction points for its binding to voltage-gated Kv1.1 channels. The NMR results indicate that HelaTx1 adopts a helix-loop-helix fold linked by two disulfide bonds without any β-sheets, resembling the molecular folding of other cysteine-stabilized helix-loop-helix (Cs α/α) scorpion toxins such as κ-hefutoxin, HeTx, and OmTx, as well as conotoxin pl14a. A series of alanine-scanning analogs revealed a broad surface on the toxin molecule largely comprising positively-charged residues that is crucial for interaction with voltage-gated Kv1.1 channels. Interestingly, the functional dyad, a key molecular determinant for activity against voltage-gated potassium channels in other toxins, is not present in HelaTx1.

siRNA-mediated Inhibition of hTERC Enhances Radiosensitivity of Cervical Cancer

  • Chen, Min;Xing, Li-Na
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.5975-5979
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    • 2012
  • Background: To investigate the influence of telomerase activity, apoptosis, radiosensitivity of cervical cancer after siRNA-mediated knockdown of telomerase RNA and evaluate in vivo growth with gene interference. Methods: We studied siRNA-targeting-telomerase RNA transfection into the Hela cell line. Expression of hTERC mRNA was detected by RT-PCR and telomerase activity was measured by the TRAP assay. Growth inhibition was determined by MTT assay and radiosensitivity of the cervical cancer cells was examined by colony formation assay. In addtion, effects of hTERC inhibition in vivo were studied by injection of siRNA-transfected Hela cells into nude mice. Results: The hTERC siRNA effectively downregulated the expression of hTERC mRNA and also reduced the telomerase activity to 30% of the untreated control vlaue. The viability of hTERC siRNA transfected Hela cells was reduced by 44.7% after transfection. After radiation treatment, the radiosensitivity of Hela cells with hTERC knockdown was increased. In vivo, the tumors developing from the hTERC siRNA-transfected cells were of reduced size, indicating that the hTERT siRNA also depressed the tumorigenic potential of the Hela cells. Conclusions: Our results supported the concept of siRNA-mediated inhibition of telomerase mRNA which could inhibit the expression of hTERC and telomerase activity. Furthermore, radiosensitivity was upregulated after knockdown the hTERC in vivo and in vitro.

Synergistic Anticancer Activity of 5-Aminolevulinic Acid Photodynamic Therapy in Combination with Low-dose Cisplatin on Hela Cells

  • Wei, Xiao-Qiang;Ma, Hui-Qing;Liu, Ai-Hong;Zhang, You-Zhong
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.3023-3028
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    • 2013
  • Objective: Photodynamic therapy (PDT ) is a promising modality for the treatment of various tumors. In order to assist in optimizing treatment, we applied 5-ALA/PDT in combination with low-dose cisplatin to evaluate cytotoxicity in Hela cells. Methods: Antiproliferative effects of 5-ALA/PDT and cisplatin, alone and in combination, were assessed using MTT assay. To examine levels of apoptosis, Hela cells treated with 5-ALA/PDT, and combination treatment were assessed with Annexin-V/PI by flow cytometry. To investigate the molecular mechanisms underlying alterations in cell proliferation and apoptosis, Western blot analysis was conducted to determine the expression of p53, p21, Bax and Bcl-2 proteins. Results: MTT assays indicated that combination treatment obviously decreased the viability of Hela cells compared to individual drug treatment. In addition, it was confirmed that exposure of Hela cells to 5-ALA/PDT in combination with low-dose cisplatin resulted in more apoptosis in vitro. Synergistic anticancer activity was related to upregulation p53 expression and alteration in expression of p21, Bcl-2 and Bax. Conclusion: Our findings suggest that administration of 5-ALA/PDT in combination with the low-dose cisplatin may be an effective and feasible therapy for cervical cancer.

Effect of Rosmarinus officinalis L. on Growth Inhibition and Apoptosis Induction in Cancer Cells (로즈마리(Rosmarinus offcinalis L.) 분획물의 암세포에 대한 성장억제 및 세포사멸 유도 효과)

  • Choi, Jun-Hyeok;Kim, Hyuk-Il;Lee, In-Seon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.8
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    • pp.1008-1015
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    • 2009
  • The goal of this study was to evaluate the anticancer effect of Rosmarinus officinalis L. In this study induction of apoptosis by methanol extract of rosemary and their fractions were investigated in vitro. In examining the effect of rosemary methanol extract on the inhibition of growth of Hela, HepG2, A549, AGS cells and HT-29 cell, it was found that the methanol extract of rosemary and their fractions demonstrated a cytotoxic effect in a dose-dependent manner; in addition, hexane and chloroform fractions showed a particularly high cytotoxic effect on Hela and AGS cells. The results showed that the hexane and chloroform fractions of rosemary have cytotoxic effect which are related to the activity of the essential oil in the rosemary. Apoptosis in Hela and AGS cells mediated by the hexane and chloroform fractions was associated with the increase of cleaved caspase-3 levels and cleaved PARP. Therefore, with more researches on identification and action mechanism of active compound, the hexane and chloroform fractions are expected to be natural sources for the developments of functional food and medical agents to prevent gastric cancer and uterus cancer.

Mechanisms of Hela Cell Apoptosis Induced by Abnormal Savda Munziq Total Phenolics Combined with Chemotherapeutic Agents

  • Zhang, Yun-Xia;Abliz, Guzalnur;Ye, Wei-Jun;Mutalipu, Zuohelaguli;Li, Xiao-Wen;Wang, Hai-Qin;Buranjiang, Gulimire;Upur, Halmurat
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.743-747
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    • 2014
  • Objective: To investigate the effects of abnormal Savda Munziq (ASMq) total phenolics combined with cisplatin and docetaxel on the Hela cell growth. Methods: In vivo cultured Hela cells were treated with cisplatin, docetaxel, total phenolics, cisplatin+total phenolics or docetaxel+total phenolics. MTT was performed to assess inhibition of cell proliferation, flow cytometry to detect apoptosis, and semi-quantitative RT-PCR to test for survivin and Bcl-2 expression. Results: The total phenolics, cisplatin and docetaxel had significant inhibitory and apoptosis-promoting effects on Hela cells (P<0.05), with the early apoptotic rates of $12.8{\pm}0.70%$, $18.9{\pm}3.79%$ and $15.8{\pm}3.8%$; the total phenolics, cisplatin and docetaxel significantly decreased the expression of Bcl-2 and survivin (all P<0.01), especially when used in combination. Conclusion: ASMq total phenolics, combined with cisplatin and docetaxel, could promote the apoptosis of Hela cells possibly through reducing the expression of Bcl-2 and survivin.

Identification of piRNAs in Hela cells by massive parallel sequencing

  • Lu, Yilu;Li, Chao;Zhang, Kun;Sun, Huaqin;Tao, Dachang;Liu, Yunqiang;Zhang, Sizong;Ma, Yongxin
    • BMB Reports
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    • v.43 no.9
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    • pp.635-641
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    • 2010
  • Piwi proteins and Piwi-interacting RNAs (piRNAs) have been implicated in transposon control in germline from Drosophila to mammals. To examine the profile of small RNA expression in human cancer cells and explore difference in small RNA transcriptome, small RNA libraries prepared from wildtype, HILI overexpressed and HILI knockdowned Hela cells were sequenced using Solexa technology. piRNAs and other repeat-associated small RNAs were observed in Hela cells. By using in situ hybridization, piR-49322 was localized in the nucleolus and around the periphery of nuclear membrane in Hela cells. Following the overexpression of HILI, the retrotransposon elements LINE1 was significantly repressed, while LINE1-associated small RNAs decreased in abundance. The present study demonstrated that HILI along with piRNAs plays a role in LINE1 suppression in Hela cancer cell line.

Cytotoxic Triterpenoids from the Fruits of Ligustrum japonicum

  • Thi Ngo, Quynh-Mai;Cao, Thao Quyen;Woo, Mi Hee;Min, Byung Sun;Weon, Kwon-Yeon
    • Natural Product Sciences
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    • v.24 no.2
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    • pp.93-98
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    • 2018
  • Medicinal plants are potential sources of anticancer agents screening. A large number of phytochemicals, including triterpenoids, have been reported to have significant cytotoxic effects on cancer cells. From the fruits of Ligustrum japonicum Thunb., thirteen triterpenoids (1 - 13) were isolated and evaluated for their cytotoxic activity against Hela and HL-60 cells. As results, 8 (oleanolic acid) showed significant effects on Hela with $IC_{50}$ values of $5.5{\mu}M$, and moderate effects on HL-60 cells with $IC_{50}$ values of $55.9{\mu}M$. Meanwhile, 10 (oleanderic acid) and 11 ($3{\beta}$-acetoxy-urs-12-en-28-oic acid) exhibited moderate inhibitory effects on Hela with $IC_{50}$ value of 55.0 and $68.8{\mu}M$, respectively. Moreover, 10 showed cytotoxic effect on HL-60 cell line with $IC_{50}$ value of $63.9{\mu}M$. To our knowledge, this is the first report that oleanderic acid was isolated from L. japonicum and investigated in cytotoxic effects on Hela and HL-60 cells.