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Ph阳性急性淋巴细胞白血病细胞株Sup-B15与基质细胞共培养后对伊马替尼敏感性的变化
OBJECTIVE: To investigate the sensitivity of imatinib (IM) on Sup-B15 Ph(+) acute lmphoblastic leukemia (ALL) cells indused by stromal cells OP9, and to further explore its mechanism. METHODS: The study is divided into two group, Sup -B15 cells group and co-cultured with OP9 cells group (Sup-B15/OP9...
Formato: | Online Artículo Texto |
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Lenguaje: | English |
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Editorial office of Chinese Journal of Hematology
2015
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7343075/ https://www.ncbi.nlm.nih.gov/pubmed/26134008 http://dx.doi.org/10.3760/cma.j.issn.0253-2727.2015.06.003 |
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collection | PubMed |
description | OBJECTIVE: To investigate the sensitivity of imatinib (IM) on Sup-B15 Ph(+) acute lmphoblastic leukemia (ALL) cells indused by stromal cells OP9, and to further explore its mechanism. METHODS: The study is divided into two group, Sup -B15 cells group and co-cultured with OP9 cells group (Sup-B15/OP9 group). The inhibitory effects of IM on leukemia cells were measured by CCK-8 test, and the apoptosis by Annexin V/7-AAD dyeing and the percentage of CD 34(+) CD38(−)leukemia cells were determined by flow cytometry. ALDH1, CD144, and β-catenin mRNA were detected by real-time RT-PCR, protein levels by Western blot. Inmunoprecipitation was used to detect the level of β-catenin connected to CD144. RESULTS: IM presented inhibitory effects on Sup-B15 and Sup-B15/OP9 cells at multiple concentrations from 10 µmol/L to 45 µmol/L. The IC(50) of IM on Sup-B15/OP and Sup-B15 cells were 35.8 µmol/L and 6.3 µmol/L, respectively (P<0.05). After 24 h of 30 µmol/L IM treatment, the percentages of apoptosis cells in Sup-B15/OP9 and Sup-B 15 cell were (14.24±2.11)% and (3.45±0.68)%, respectively (P<0.05). The percentage of CD34(+)CD38(−)cells in Sup-B15/OP9 group was significantly higher than that in Sup-B15 group [(3.42±0.28)% vs (0.16±0.15)%, P<0.05]. As compared to Sup-B15 cells, the transcription of ALDH1 in Sup-B15/OP9 group was remarkably upregulated (0.097±0.012 vs 0.046±0.010, P<0.05), and the CD133 protein level was also upregulated in Sup-B15/OP9 group. The transcription of CD144 in Sup-B15/OP9 group was remarkably upregulated compared with Sup-B15 group (0.103±0.015 vs 0.010±0.003, P<0.05), as well as the CD144 protein. β-catenin mRNA transcription has no obvious changes between Sup-B15 group and Sup-B15/OP9 group (P>0.05), while the whole β-catenin protein and the cell nucleus β-catenin significantly increased, as well as the β-catenin protein combined with CD144. CONCLUSION: Co-cultured with OP9 cells, Sup-B15 cells show less sensitivity to imatinib. The raising activity of CD144 and CD144/β-catenin signaling may work in this procession. |
format | Online Article Text |
id | pubmed-7343075 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Editorial office of Chinese Journal of Hematology |
record_format | MEDLINE/PubMed |
spelling | pubmed-73430752020-07-16 Ph阳性急性淋巴细胞白血病细胞株Sup-B15与基质细胞共培养后对伊马替尼敏感性的变化 Zhonghua Xue Ye Xue Za Zhi 论著 OBJECTIVE: To investigate the sensitivity of imatinib (IM) on Sup-B15 Ph(+) acute lmphoblastic leukemia (ALL) cells indused by stromal cells OP9, and to further explore its mechanism. METHODS: The study is divided into two group, Sup -B15 cells group and co-cultured with OP9 cells group (Sup-B15/OP9 group). The inhibitory effects of IM on leukemia cells were measured by CCK-8 test, and the apoptosis by Annexin V/7-AAD dyeing and the percentage of CD 34(+) CD38(−)leukemia cells were determined by flow cytometry. ALDH1, CD144, and β-catenin mRNA were detected by real-time RT-PCR, protein levels by Western blot. Inmunoprecipitation was used to detect the level of β-catenin connected to CD144. RESULTS: IM presented inhibitory effects on Sup-B15 and Sup-B15/OP9 cells at multiple concentrations from 10 µmol/L to 45 µmol/L. The IC(50) of IM on Sup-B15/OP and Sup-B15 cells were 35.8 µmol/L and 6.3 µmol/L, respectively (P<0.05). After 24 h of 30 µmol/L IM treatment, the percentages of apoptosis cells in Sup-B15/OP9 and Sup-B 15 cell were (14.24±2.11)% and (3.45±0.68)%, respectively (P<0.05). The percentage of CD34(+)CD38(−)cells in Sup-B15/OP9 group was significantly higher than that in Sup-B15 group [(3.42±0.28)% vs (0.16±0.15)%, P<0.05]. As compared to Sup-B15 cells, the transcription of ALDH1 in Sup-B15/OP9 group was remarkably upregulated (0.097±0.012 vs 0.046±0.010, P<0.05), and the CD133 protein level was also upregulated in Sup-B15/OP9 group. The transcription of CD144 in Sup-B15/OP9 group was remarkably upregulated compared with Sup-B15 group (0.103±0.015 vs 0.010±0.003, P<0.05), as well as the CD144 protein. β-catenin mRNA transcription has no obvious changes between Sup-B15 group and Sup-B15/OP9 group (P>0.05), while the whole β-catenin protein and the cell nucleus β-catenin significantly increased, as well as the β-catenin protein combined with CD144. CONCLUSION: Co-cultured with OP9 cells, Sup-B15 cells show less sensitivity to imatinib. The raising activity of CD144 and CD144/β-catenin signaling may work in this procession. Editorial office of Chinese Journal of Hematology 2015-06 /pmc/articles/PMC7343075/ /pubmed/26134008 http://dx.doi.org/10.3760/cma.j.issn.0253-2727.2015.06.003 Text en 2015年版权归中华医学会所有 http://creativecommons.org/licenses/by-nc-sa/3.0/ This work is licensed under a Creative Commons Attribution 3.0 License (CC-BY-NC). The Copyright own by Publisher. Without authorization, shall not reprint, except this publication article, shall not use this publication format design. Unless otherwise stated, all articles published in this journal do not represent the views of the Chinese Medical Association or the editorial board of this journal. |
spellingShingle | 论著 Ph阳性急性淋巴细胞白血病细胞株Sup-B15与基质细胞共培养后对伊马替尼敏感性的变化 |
title | Ph阳性急性淋巴细胞白血病细胞株Sup-B15与基质细胞共培养后对伊马替尼敏感性的变化 |
title_full | Ph阳性急性淋巴细胞白血病细胞株Sup-B15与基质细胞共培养后对伊马替尼敏感性的变化 |
title_fullStr | Ph阳性急性淋巴细胞白血病细胞株Sup-B15与基质细胞共培养后对伊马替尼敏感性的变化 |
title_full_unstemmed | Ph阳性急性淋巴细胞白血病细胞株Sup-B15与基质细胞共培养后对伊马替尼敏感性的变化 |
title_short | Ph阳性急性淋巴细胞白血病细胞株Sup-B15与基质细胞共培养后对伊马替尼敏感性的变化 |
title_sort | ph阳性急性淋巴细胞白血病细胞株sup-b15与基质细胞共培养后对伊马替尼敏感性的变化 |
topic | 论著 |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7343075/ https://www.ncbi.nlm.nih.gov/pubmed/26134008 http://dx.doi.org/10.3760/cma.j.issn.0253-2727.2015.06.003 |
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