Abstract
Multidrug resistance is one of the major reasons colorectal cancer (CRC) chemotherapy-based treatments fail, and novel biologically based therapies are urgently needed. Src homology 3 (SH3)-domain GRB2-like protein 1 (SH3GL1) is a membrane-bound protein which was found to be involved in tumor formation, progression, and metastasis. In this study, immunohistochemistry staining, quantitative real-time polymerase chain reaction (qRT-PCR), and western blot analysis revealed a high expression of SH3GL1 in human CRC tumor specimens and several CRC cells resistant to chemotherapeutics. Cell Counting Kit-8 (CCK-8) assay showed that transfection of pCDNA3.1(+)-SH3GL1 increased while transfection of SH3GL1 siRNA decreased cell viability in response to 5-fluorouracil (5-FU) treatment (P < 0.05). Further studies indicated that transfection of SH3GL1 siRNA significantly downregulated multidrug resistance protein 1 (MDR1)/P-glycoprotein expression (P < 0.05), decreased MDR1 promoter activity and activator protein-1 (AP-1) binding activity (P < 0.05), and inhibited the activation of epidermal growth factor receptor (EGFR) and extracellular signal-regulated kinases 1/2 (ERK1/2) signaling (P < 0.05) in CRC cells resistant to chemotherapeutics. Transfection of pCDNA3.1(+)-SH3GL1 caused the opposite effect. Additionally, pre-treatment with either EGFR kinase inhibitor PD153035 or ERK1/2 kinase inhibitor PD98059 in HCT116/5-FU cells partly inhibits P-glycoprotein expression and AP-1 binding activity (P < 0.05). In conclusion, we confirmed that inhibition of SH3GL1 reverses multidrug resistance through declining P-glycoprotein expression via the EGFR/ERK/AP-1 pathway.
Similar content being viewed by others
References
Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin. 2011;61(2):69–90.
Kou C, Zhou T, Han X, Zhuang H, Qian H. Lrig1, a 3p tumor suppressor, represses egfr signaling and is a novel epigenetic silenced gene in colorectal cancer. Biochem Biophys Res Commun. 2015;464(2):519–25.
Li Q, Sui H. Abstract 2328: COX-2 induce mediated multidrug resistance by activating jnk signal transduction pathway in colorectal cancer. Cancer Res. 2014;74(19 Supplement):2328.
Callaghan R, Luk F, Bebawy M. Inhibition of the multidrug resistance P-glycoprotein: time for a change of strategy? Drug Metab Dispos. 2014;42(4):623–31.
Gottesman MM, Pastan IH. The role of multidrug resistance efflux pumps in cancer: revisiting a JNCI publication exploring expression of the mdr1 (P-glycoprotein) gene. J Natl Cancer Inst. 2015;107(9). doi:10.1093/jnci/djv222.
Wang MQ, Kim W, Gao G, Torrey TA, Morse 3rd HC, De Camilli P, et al. Endophilins interact with Moloney murine leukemia virus Gag and modulate virion production. J Biol. 2003;3(1):4. doi:10.1186/1475-4924-3-4.
Yam JW, Jin DY, So CW, Chan LC. Identification and characterization of EBP, a novel EEN binding protein that inhibits Ras signaling and is recruited into the nucleus by the MLL-EEN fusion protein. Blood. 2004;103(4):1445–53.
Matsutani T, Hiwasa T, Takiguchi M, Oide T, Kunimatsu M, Saeki N, et al. Autologous antibody to src-homology 3-domain GRB2-like 1 specifically increases in the sera of patients with low-grade gliomas. J Exp Clin Cancer Res. 2012;31:85. doi:10.1186/1756-9966-31-85.
Zhao L, Wang H, Liu C, Liu Y, Wang X, Wang S, et al. Promotion of colorectal cancer growth and metastasis by the LIM and SH3 domain protein 1. Gut. 2010;59(9):1226–35.
Slupianek A, Schmutte C, Tombline G, Nieborowska-Skorska M, Hoser G, Nowicki MO, et al. BCR/ABL regulates mammalian RecA homologs, resulting in drug resistance. Mol Cell. 2001;8(4):795–806.
Yang LZ, Zhang B, Kou WZ, Niu HR, Xue HC, Li XR. The clinical implication of SH3GL1 expression in paclitaxel-resistant breast cancer. Chin J Biochem Mol Biol. 2015;31(10):1064–70.
Lua BL, Low BC. Activation of EGF receptor endocytosis and ERK1/2 signaling by BPGAP1 requires direct interaction with EEN/endophilin II and a functional RhoGAP domain. J Cell Sci. 2005;118(Pt 12):2707–21.
Ayuso-Sacido A, Moliterno JA, Kratovac S, Kapoor GS, O’Rourke DM, Holland EC, et al. Activated EGFR signaling increases proliferation, survival, and migration and blocks neuronal differentiation in post-natal neural stem cells. J Neuro-Oncol. 2010;97(3):323–37.
Mendelsohn J, Prewett M, Rockwell P, Goldstein NI. CCR 20th anniversary commentary: a chimeric antibody, C225, inhibits EGFR activation and tumor growth. Clin Cancer Res. 2015;21(2):227–9.
Nautiyal J, Du J, Yu Y, Kanwar SS, Levi E, Majumdar AP. EGFR regulation of colon cancer stem-like cells during aging and in response to the colonic carcinogen dimethylhydrazine. Am J Physiol Gastrointest Liver Physiol. 2012;302(7):G655–63.
Munoz JL, Rodriguez-Cruz V, Greco SJ, Nagula V, Scotto KW, Rameshwar P. Temozolomide induces the production of epidermal growth factor to regulate MDR1 expression in glioblastoma cells. Mol Cancer Ther. 2014;13(10):2399–411.
Uchibori K, Kasamatsu A, Sunaga M, Yokota S, Sakurada T, Kobayashi E, et al. Establishment and characterization of two 5-fluorouracil-resistant hepatocellular carcinoma cell lines. Int J Oncol. 2012;40(4):1005–10.
Huang EW, Xue SJ, Li XY, Xu SW, Cheng JD, Zheng JX, et al. EEN regulates the proliferation and survival of multiple myeloma cells by potentiating IGF-1 secretion. Biochem Biophys Res Commun. 2014;447(2):271–7.
Baldassarre T, Watt K, Truesdell P, Schneider M, Sengupta S, Craig AW. Defining molecular mechanisms linking endophilin A2 to metastasis in human breast cancer models. Cancer Res. 2015;75(15 Supplement):2260.
Wu Q, Yang Z, Nie Y, Shi Y, Fan D. Multi-drug resistance in cancer chemotherapeutics: mechanisms and lab approaches. Cancer Lett. 2014;347(2):159–66.
Soto-Guzman A, Robledo T, Lopez-Perez M, Salazar EP. Oleic acid induces ERK1/2 activation and AP-1 DNA binding activity through a mechanism involving Src kinase and EGFR transactivation in breast cancer cells. Mol Cell Endocrinol. 2008;294(1–2):81–91.
Feng D, Ma Y, Liu J, Xu L, Zhang Y, Qu J, et al. Cbl-b enhances sensitivity to 5-fluorouracil via EGFR- and mitochondria-mediated pathways in gastric cancer cells. Int J Mol Sci. 2013;14(12):24399–411.
Vehlow A, Soong D, Vizcay-Barrena G, Bodo C, Law AL, Perera U, et al. Endophilin, lamellipodin, and mena cooperate to regulate F-actin-dependent EGF-receptor endocytosis. EMBO J. 2013;32(20):2722–34.
Renard HF, Simunovic M, Lemiere J, Boucrot E, Garcia-Castillo MD, Arumugam S, et al. Endophilin-A2 functions in membrane scission in clathrin-independent endocytosis. Nature. 2015;517(7535):493–6.
Mitra SK, Schlaepfer DD. Integrin-regulated FAK-Src signaling in normal and cancer cells. Curr Opin Cell Biol. 2006;18(5):516–23.
Zhang J, Fan J, Tian Q, Song Z, Zhang JF, Chen Y. Characterization of two distinct modes of endophilin in clathrin-mediated endocytosis. Cell Signal. 2012;24(11):2043–50.
Wu X, Gan B, Yoo Y, Guan JL. FAK-mediated src phosphorylation of endophilin A2 inhibits endocytosis of MT1-MMP and promotes ECM degradation. Dev Cell. 2005;9(2):185–96.
Ross JA, Chen Y, Muller J, Barylko B, Wang L, Banks HB, et al. Dimeric endophilin A2 stimulates assembly and GTPase activity of dynamin 2. Biophys J. 2011;100(3):729–37.
Baldassarre T, Watt K, Truesdell P, Meens J, Schneider MM, Sengupta SK, et al. Endophilin A2 promotes TNBC cell invasion and tumor metastasis. Mol Cancer Res. 2015;13(6):1044–55.
Chen J, Ding Z, Peng Y, Pan F, Li J, Zou L, et al. HIF-1alpha inhibition reverses multidrug resistance in colon cancer cells via downregulation of MDR1/P-glycoprotein. PLoS One. 2014;9(6):e98882.
Sui H, Zhou S, Wang Y, Liu X, Zhou L, Yin P, et al. COX-2 contributes to P-glycoprotein-mediated multidrug resistance via phosphorylation of c-Jun at Ser63/73 in colorectal cancer. Carcinogenesis. 2011;32(5):667–75.
Chen Q, Bian Y, Zeng S. Involvement of AP-1 and NF-kappaB in the up-regulation of P-gp in vinblastine resistant Caco-2 cells. Drug Metab Pharmacokinet. 2014;29(2):223–6.
Daschner PJ, Ciolino HP, Plouzek CA, Yeh GC. Increased AP-1 activity in drug resistant human breast cancer MCF-7 cells. Breast Cancer Res Treat. 1999;53(3):229–40.
Han L, Wang Y, Guo X, Zhou Y, Zhang J, Wang N, et al. Downregulation of MDR1 gene by cepharanthine hydrochloride is related to the activation of c-Jun/JNK in K562/ADR cells. Biomed Res Int. 2014;2014:164391. doi:10.1155/2014/164391.
Shi B, Isseroff RR. Epidermal growth factor (EGF)-mediated DNA-binding activity of AP-1 is attenuated in senescent human epidermal keratinocytes. Exp Dermatol. 2005;14(7):519–27.
Ung CY, Li H, Ma XH, Jia J, Li BW, Low BC, et al. Simulation of the regulation of EGFR endocytosis and EGFR-ERK signaling by endophilin-mediated RhoA-EGFR crosstalk. FEBS Lett. 2008;582(15):2283–90.
X-H T, Deng S, Li M, Lu M-S. Cross-reacting material 197 reverses the resistance to paclitaxel in paclitaxel-resistant human ovarian cancer. Tumor Biol. 2015;1–8.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
The study was performed with the approval of the Ethics Committee of the Second Affiliated Hospital of Xi’an Jiaotong University. Written informed consents were obtained from each patient before study initiation.
Conflicts of interest
None.
Rights and permissions
About this article
Cite this article
Guan, H., Zhao, P., Dai, Z. et al. SH3GL1 inhibition reverses multidrug resistance in colorectal cancer cells by downregulation of MDR1/P-glycoprotein via EGFR/ERK/AP-1 pathway. Tumor Biol. 37, 12153–12160 (2016). https://doi.org/10.1007/s13277-016-5092-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13277-016-5092-0