浏览全部资源
扫码关注微信
HeLa细胞上调Beclin1/自噬通路对多柔比星治疗抵抗
HeLa cells up-regulate Beclin1/autophagy pathway to confer resistance to doxorubicin
- 第三军医大学学报 2019年41卷第13期 页码：1216-1221
- 作者机构：
1.100049 北京，航天中心医院妇产科
2.100048 北京，解放军总医院第六医学中心
- 作者简介：
[郭艳军，E-mail: gyj_w@126.com GUO Yanjun, E-mail: gyj_w@126.com
- 基金信息：
DOI：10.16016/j.1000-5404.201903084
中图分类号： R737.33;R966;R979.14收稿：2019-03-10，
修回：2019-05-01，
纸质出版：2019-07-15
- 稿件说明：
移动端阅览
郭艳军, 王淑为. HeLa细胞上调Beclin1/自噬通路对多柔比星治疗抵抗[J]. 第三军医大学学报, 2019,41(13):1216-1221.
Yanjun GUO, Shuwei WANG. HeLa cells up-regulate Beclin1/autophagy pathway to confer resistance to doxorubicin[J]. Journal of Third Military Medical University, 2019, 41(13): 1216-1221.
郭艳军, 王淑为. HeLa细胞上调Beclin1/自噬通路对多柔比星治疗抵抗[J]. 第三军医大学学报, 2019,41(13):1216-1221. DOI： 10.16016/j.1000-5404.201903084.
Yanjun GUO, Shuwei WANG. HeLa cells up-regulate Beclin1/autophagy pathway to confer resistance to doxorubicin[J]. Journal of Third Military Medical University, 2019, 41(13): 1216-1221. DOI： 10.16016/j.1000-5404.201903084.
摘要
目的
2
观察多柔比星(doxorubicin
DOX)在HeLa细胞中诱导保护性自噬反应的作用，探讨HeLa细胞对DOX治疗抵抗的可能机制。
方法
2
用不同剂量(0、2、4、8、16 mg/L)多柔比星处理人宫颈癌HeLa细胞，CCK-8法检测细胞活力，定量PCR检测DOX处理对自噬相关基因mRNA的影响，Western blot分析DOX处理对自噬相关蛋白表达的影响，用siRNA沉默Beclin1，CCK-8法检测DOX对HeLa细胞活力的影响，把含有Beclin1启动子片段的报告质粒转入HeLa细胞，通过报告基因系统检测DOX处理对Beclin1启动子活性的影响，用转录抑制剂放线菌素D预处理HeLa细胞，然后用定量PCR分析DOX处理不同时间对Beclin1 mRNA稳定性的影响。
结果
2
DOX可剂量依赖性抑制HeLa细胞活力，并诱导细胞的保护性自噬反应，抑制自噬可增强HeLa细胞对DOX的敏感性。DOX可特异性地升高HeLa细胞中Beclin1的mRNA和蛋白水平(
P
<
0.05)。沉默Beclin1可增强DOX对HeLa细胞的杀伤效果。DOX处理不影响HeLa细胞中Beclin1的启动子活性(
P
>
0.05)，但显著增强其mRNA的稳定性(
P
<
0.05)。
结论
2
DOX可能通过增强Beclin1/自噬通路而导致HeLa细胞对其耐受。
Abstract
Objective
2
To investigate the role of doxorubicin (DOX) in the induction of protective autophagy in HeLa cells and explore the possible mechanism of DOX resistance.
Methods
2
HeLa cells were treated with DOX at different doses (0
2
4
8 and 16 mg/L) for 24 h
and the cell viability was measured by CCK-8 assay. The mRNA levels of autophagy-related genes were assessed using qRT-PCR
and the protein levels were detected by Western blotting. After Beclin1 was knockdowned through siRNA
the effect of DOX on cell viability was detected by CCK-8 assay. The effect of DOX on the promoter activity of Beclin1 in HeLa cells was assessed using the reporter gene assay after transfection with a plasmid containing Beclin1 promoter. The stability of Beclin1 mRNA was analyzed with qPCR in HeLa cells after treatment with DOX alone or DOX combined with the transcription inhibitor actinomycin D.
Results
2
DOX inhibited the cell viability in HeLa cells in a dose-dependent manner. DOX also induced protective autophagy in HeLa cells. Inhibition of autophagy enhanced the sensitivity of HeLa cells to DOX. DOX treatment resulted in special up-regulation of Beclin1 at both mRNA and protein levels in HeLa ce
lls (
P
<
0.05). Knockdown of Beclin1 significantly increased DOX-triggered proliferation suppression in HeLa cells. DOX treatment did not significantly affect the promoter activity of Beclin1
but obviously lowered the mRNA stability of Beclin1 in HeLa cells.
Conclusion
2
DOX may induce drug resistance in HeLa cells through enhancing the Beclin1/autophagy pathway.
关键词
Keywords
references
BRAY F, FERLAY J, SOERJOMATARAM I, et al . Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J ] . CA Cancer J Clin, 2018, 68(6): 394-424. DOI:10.3322/caac.21492.
COHEN P A, JHINGRAN A, OAKNIN A, et al . Cervical cancer[J ] . Lancet, 2019, 393(10167): 169-182. DOI:10.1016/S0140-6736(18)32470-X.
RIVANKAR S. An overview of doxorubicin formulations in cancer therapy[J]. J Cancer Res Ther, 2014, 10(4): 853-858. DOI:10.4103/0973-1482.139267.
MOHAJERI M, SAHEBKAR A. Protective effects of curcumin against doxorubicin-induced toxicity and resistance: A review[J]. Crit Rev Oncol Hematol, 2018, 122: 30-51. DOI:10.1016/j.critrevonc.2017.12.005.
SAHA S, PANIGRAHI D P, PATIL S, et al . Autophagy in health and disease: A comprehensive review[J ] . Biomed Pharmacother, 2018, 104: 485-495. DOI:10.1016/j.biopha.2018.05.007.
PRIETO-DOMÍNGUEZ N, GARCIA-MEDIAVILLA M V, SANCHEZ-CAMPOS S, et al . Autophagy as a molecular target of flavonoids underlying their protective effects in human disease[J ] . Curr Med Chem, 2018, 25(7): 814-838. DOI:10.2174/0929867324666170918125155.
TAYLOR M A, DAS B C, RAY S K. Targeting autophagy for combating chemoresistance and radioresistance in glioblastoma[J]. Apoptosis, 2018, 23(11/12): 563-575. DOI:10.1007/s10495-018-1480-9.
YUNIARTI L, MUSTOFA M, ARYANDONO T, et al . Synergistic action of 1, 2-epoxy-3 (3- (3, 4-dimethoxyphenyl)- 4H-1-benzopiyran-4-on) propane with doxorubicin and cisplatin through increasing of p53, TIMP-3, and microRNA-34a in cervical cancer cell line (HeLa)[J ] . Asian Pac J Cancer Prev, 2018, 19(10): 2955-2962. DOI:10.22034/APJCP.2018.19.10.2955.
XIA Y, XU T, ZHAO M, et al . Delivery of doxorubicin for human cervical carcinoma targeting therapy by folic acid-modified selenium nanoparticles[J ] . Int J Mol Sci, 2018, 19(11): E3582. DOI:10.3390/ijms19113582.
QIU F, ZHAO X. Study on the sensitivity of primary tumor cells of patients with cervical cancer to chemotherapeutic combinations[J]. J BUON, 2018, 23(1): 117-123.
MUTLU P, YALÇIN AZARKAN S, TAGHAVI POURIANAZAR N, et al . Determination of the relationship between doxorubicin resistance and Wnt signaling pathway in HeLa and K562 cell lines[J ] . EXCLI J, 2018, 17: 386-398. DOI:10.17179/excli2018-1129.
WAGNER W, KANIA K D, BLAUZ A, et al . The lactate receptor (HCAR1/GPR81) contributes to doxorubicin chemoresistance via ABCB1 transporter up-regulation in human cervical cancer HeLa cells[J ] . J Physiol Pharmacol, 2017, 68(4): 555-564..
XU J, PAN Q, JU W.Ras inhibition by zoledronic acid effectively sensitizes cervical cancer to chemotherapy[J].Anticancer Drugs, 2019..DOI: 10.1097/CAD.0000000000000779.
GUAN S, LU J X, ZHAO Y L, et al . TAK1 inhibitor 5Z-7-oxozeaenol sensitizes cervical cancer to doxorubicin-induced apoptosis[J ] . Oncotarget, 2017, 8(20): 33666-33675. DOI:10.18632/oncotarget.16895.
GUO K Y, HAN L L, LI X Y, et al . Novel proteasome inhibitor delanzomib sensitizes cervical cancer cells to doxorubicin-induced apoptosis via stabilizing tumor suppressor proteins in the p53 pathway[J ] . Oncotarget, 2017, 8(69): 114123-114135. DOI:10.18632/oncotarget.23166..
DAGLIOGLU C, KACI F N. Cascade therapy with doxorubicin and survivin-targeted tailored nanoparticles: An effective alternative for sensitization of cancer cells to chemotherapy[J]. Int J Pharm, 2019, 561: 74-81. DOI:10.1016/j.ijpharm.2019.02.036.
WU Y R, NI Z H, YAN X J, et al . Targeting the MIR34C-5p-ATG4B-autophagy axis enhances the sensitivity of cervical cancer cells to pirarubicin[J ] . Autophagy, 2016, 12(7): 1105-1117. DOI:10.1080/15548627.2016.1173798.
XIONG H, NI Z, HE J, et al . LncRNA HULC triggers autophagy via stabilizing Sirt1 and attenuates the chemosensitivity of HCC cells[J ] . Oncogene, 2017, 36(25): 3528-3540. D OI:10.1038/onc.2016.521..
KRANZBÜHLER B, SALEMI S, MORTEZAVI A, et al . Combined N-terminal androgen receptor and autophagy inhibition increases the antitumor effect in enzalutamide sensitive and enzalutamide resistant prostate cancer cells[J ] . Prostate, 2019, 79(2):206-214. DOI: 10.1002/pros.23725.
SUN W L, WANG L, LUO J, et al . Ambra1 modulates the sensitivity of breast cancer cells to epirubicin by regulating autophagy via ATG12[J ] . Cancer Sci, 2018, 109(10): 3129-3138. DOI:10.1111/cas.13743.
ZHANG N, WU Y R, LYU X L, et al . HSF1 upregulates ATG4B expression and enhances epirubicin-induced protective autophagy in hepatocellular carcinoma cells[J ] . Cancer Lett, 2017, 409: 81-90. DOI:10.1016/j.canlet.2017.08.039.
CHEN C, LU L, YAN S C, et al . Autophagy and doxorubicin resistance in cancer[J ] . Anticancer Drugs, 2018, 29(1): 1-9. DOI:10.1097/CAD.0000000000000572.
FERNANDES Â, AZEVEDO M M, PEREIRA O, et al . Proteolytic systems and AMP-activated protein kinase are critical targets of acute myeloid leukemia therapeutic approaches[J ] . Onc otarget, 2015, 6(31): 31428-31440. DOI:10.18632/oncotarget.2947.
DI FAZIO P, MATROOD S. Targeting autophagy in liver cancer[J]. Transl Gastroenterol Hepatol, 2018, 3: 39. DOI:10.21037/tgh.2018.06.09.
WU C L, LIU J F, LIU Y, et al . Beclin1 inhibition enhances paclitaxel-mediated cytotoxicity in breast cancer in vitro and in vivo [J ] . Int J Mol Med, 2019, 43(4): 1866-1878. DOI:10.3892/ijmm.2019.4089..
GANDHI M, CAUDRON-HERGER M, DIEDERICHS S. RNA motifs and combinatorial prediction of interactions, stability and localization of noncoding RNAs[J]. Nat Struct Mol Biol, 2018, 25(12): 1070-1076. DOI:10.1038/s41594-018-0155-0.
0
浏览量
0
下载量
0
CSCD
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies
