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PU-H71对增加宫颈癌细胞辐射敏感性的研究
王建凯1, 缪国英1, 王文娟2, 胡永果1, 蔡宏懿1
1甘肃省人民医院放疗科,兰州 730000; 2甘肃省第三人民医院体检中心,兰州 730000
Study of the effect of PU-H71 on increasing radiosensitivity of cervical cancer cells
Wang Jiankai1, Miao Guoying1, Wang Wenjuan2, Hu Yongguo1, Cai Hongyi1
1Department of Radiation Oncology,Gansu Provincial Hospital, Lanzhou 730000, China; 2Physical Examination Center, Gansu Third People's Hospital, Lanzhou 730000, China
Abstract：Objective To investigate the effect of heat shock protein 90 (Hsp90) inhibitor PU-H71 combined with X-ray on radioresistant human cervical cancer cells. Methods The expression levels of Hsp90 gene between cervical cancer tissues and adjacent tissues were analyzed by bioinformatics. Radioresistant cervical cancer cell lines HeLa RR and SiHa RR were obtained by fractional irradiations (2 Gy per fraction, 30 fractions). The cell lines were divided into the control group (treated with dimethyl sulfoxide), irradiation alone group, PU-H71 group (treated with 0.5 μmol/L PU-H71), and PU-H71+irradiation group (irradiation at 24 h after treatment with 0.5 μmol/L PU-H71). Cell survival was detected by clonal formation assay. Immunofluorescence assay was used to detect γH2AX foci at 1, 6, and 24 h after cell treatment. The expression level of Rad51 protein at 1, 2, 6, 12, and 24 h after cell treatment was detected using Western blot. The expression level of phosphorylated DNA-dependent protein kinase catalytic subunit (p-DNA-PKcs) was measured at 2 h after cell treatment. Cell apoptosis at 48 h after cell treatment was assessed by flow cytometry. Results PU-H71 enhanced the sensitivity of radioresistant cervical cancer cells to X-ray. Compared with the irradiation alone group, the radiation sensitization ratios (SER) of HeLa RR and SiHa RR cells at 10% survival were 1.36 and 1.27, and the apoptosis rates were increased by approximately 72.1% and 63.1% in the PU-H71+irradiation group, respectively. PU-H71 delayed the duration of γH2AX foci induced by X-ray, inhibited the phosphorylation of DNA-dependent protein kinase catalytic subunit (DNA-PKcs), thus preventing non-homologous end joining (NHEJ) repair and delaying homologous recombination repair. Conclusion PU-H71 increases the radiosensitivity of radioresistant cervical cancer cells by inhibiting the repair pathway of DNA double-strand break, which is expected to be a radiosensitizer to enhance the efficacy of radiotherapy for cervical cancer.
Wang Jiankai,Miao Guoying,Wang Wenjuan et al. Study of the effect of PU-H71 on increasing radiosensitivity of cervical cancer cells[J]. Chinese Journal of Radiation Oncology, 2023, 32(6): 551-556.
[1] Siegel RL, Miller KD, Jemal A.Cancer statistics, 2020[J]. CA Cancer J Clin, 2020,70(1):7-30. DOI: 10.3322/caac.21590. [2] de Martel C, Plummer M, Vignat J, et al. Worldwide burden of cancer attributable to HPV by site, country and HPV type[J]. Int J Cancer, 2017,141(4):664-670. DOI: 10.1002/ijc.30716. [3] Shah SC, Kayamba V, Peek RM, et al.Cancer control in low- and middle-income countries: is it time to consider screening?[J]. J Glob Oncol, 2019,5:1-8. DOI: 10.1200/JGO.18.00200. [4] Chino J, Annunziata CM, Beriwal S, et al.Radiation therapy for cervical cancer: executive summary of an ASTRO clinical practice guideline[J]. Pract Radiat Oncol, 2020,10(4):220-234. DOI: 10.1016/j.prro.2020.04.002. [5] Li Q, Wei X, Zhou ZW, et al.GADD45α sensitizes cervical cancer cells to radiotherapy via increasing cytoplasmic APE1 level[J]. Cell Death Dis, 2018,9(5):524. DOI: 10.1038/s41419-018-0452-x. [6] Hoter A, El-Sabban ME, Naim HY. The HSP90 family: structure, regulation, function,implications in health and disease[J]. Int J Mol Sci, 2018,19(9)DOI: 10.3390/ijms19092560. [7] Birbo B, Madu EE, Madu CO, et al. Role of HSP90 in Cancer[J]. Int J Mol Sci, 2021,22(19)DOI: 10.3390/ijms221910317. [8] Song Q, Wen JY, Li WP, et al.HSP90 promotes radioresistance of cervical cancer cells via reducing FBXO6-mediated CD147 polyubiquitination[J]. Cancer Sci, 2022,113(4):1463-1474. DOI: 10.1111/cas.15269. [9] Jackson SE.Hsp90: structure and function[J]. Top Curr Chem, 2013,328:155-240. DOI: 10.1007/128_2012_356. [10] Usmani SZ, Bona RD, Chiosis G, et al.The anti-myeloma activity of a novel purine scaffold HSP90 inhibitor PU-H71 is via inhibition of both HSP90A and HSP90B1[J]. J Hematol Oncol, 2010,3:40. DOI: 10.1186/1756-8722-3-40. [11] He H, Zatorska D, Kim J, et al.Identification of potent water soluble purine-scaffold inhibitors of the heat shock protein 90[J]. J Med Chem, 2006,49(1):381-390. DOI: 10.1021/jm0508078. [12] Kale Ş, Korcum AF, Dündar E, et al.HSP90 inhibitor PU-H71 increases radiosensitivity of breast cancer cells metastasized to visceral organs and alters the levels of inflammatory mediators[J]. Naunyn Schmiedebergs Arch Pharmacol, 2020,393(2):253-262. DOI: 10.1007/s00210-019-01725-z. [13] Moulick K, Ahn JH, Zong H, et al.Affinity-based proteomics reveal cancer-specific networks coordinated by Hsp90[J]. Nat Chem Biol, 2011,7(11):818-826. DOI: 10.1038/nchembio.670. [14] Liu XX, Sun C, Jin XD, et al.Genistein enhances the radiosensitivity of breast cancer cells via G?/M cell cycle arrest and apoptosis[J]. Molecules, 2013,18(11):13200-13217. DOI: 10.3390/molecules181113200. [15] Hu SL, Tan JQ, Qin LX, et al.Molecular chaperones and Parkinson's disease[J]. Neurobiol Dis, 2021,160:105527. DOI: 10.1016/j.nbd.2021.105527. [16] Nagaraju GP, Zakka KM, Landry JC, et al.Inhibition of HSP90 overcomes resistance to chemotherapy and radiotherapy in pancreatic cancer[J]. Int J Cancer, 2019,145(6):1529-1537. DOI: 10.1002/ijc.32227. [17] Modi S, Stopeck AT, Gordon MS, et al.Combination of trastuzumab and tanespimycin (17-AAG, KOS-953) is safe and active in trastuzumab-refractory HER-2 overexpressing breast cancer: a phase I dose-escalation study[J]. J Clin Oncol, 2007,25(34):5410-5417. DOI: 10.1200/JCO.2007.11.7960. [18] Youssef ME, Cavalu S, Hasan AM, et al.Role of ganetespib, an HSP90 inhibitor, in cancer therapy: from molecular mechanisms to clinical practice[J]. Int J Mol Sci, 2023, 24(5):5014. DOI:10.3390/ijms24055014. [19] Antolin AA, Clarke PA, Collins I, et al. Evolution of kinase polypharmacology across HSP90 drug discovery[J]. Cell Chem Biol, 2021,28(10):1433-1445.e3. DOI: 10.1016/j.chembiol.2021.05.004. [20] Butler LM, Ferraldeschi R, Armstrong HK, et al.Maximizing the therapeutic potential of HSP90 inhibitors[J]. Mol Cancer Res, 2015,13(11):1445-1451. DOI: 10.1158/1541-7786.MCR-15-0234. [21] Caldas-Lopes E, Cerchietti L, Ahn JH, et al.Hsp90 inhibitor PU-H71, a multimodal inhibitor of malignancy, induces complete responses in triple-negative breast cancer models[J]. Proc Natl Acad Sci U S A, 2009,106(20):8368-8373. DOI: 10.1073/pnas.0903392106. [22] Gallerne C, Prola A, Lemaire C.Hsp90 inhibition by PU-H71 induces apoptosis through endoplasmic reticulum stress and mitochondrial pathway in cancer cells and overcomes the resistance conferred by Bcl-2[J]. Biochim Biophys Acta, 2013,1833(6):1356-1366. DOI: 10.1016/j.bbamcr.2013.02.014. [23] Chiosis G, Kang Y, Sun W.Discovery and development of purine-scaffold Hsp90 inhibitors[J]. Expert Opin Drug Discov, 2008,3(1):99-114. DOI: 10.1517/17460441.3.1.99. [24] Demeyer A, Benhelli-Mokrani H, Chénais B, et al.Inhibiting homologous recombination by targeting RAD51 protein[J]. Biochim Biophys Acta Rev Cancer, 2021,1876(2):188597. DOI: 10.1016/j.bbcan.2021. 188597. [25] Feng W, Smith CM, Simpson DA, et al.Targeting non-homologous and alternative end joining repair to enhance cancer radiosensitivity[J]. Semin Radiat Oncol, 2022,32(1):29-41. DOI: 10.1016/j.semradonc.2021.09.007. [26] Liu XX, Li P, Hirayama R, et al.Genistein sensitizes glioblastoma cells to carbon ions via inhibiting DNA-PKcs phosphorylation and subsequently repressing NHEJ and delaying HR repair pathways[J]. Radiother Oncol, 2018,129(1):84-94. DOI: 10.1016/j.radonc.2018.04.005.
2023, Vol. 32 
