Doxorubicin-resistant MCF-7 Cells Research Articles

Effectiveness of Pheophorbide-a mediated Photodynamic Therapy against Doxorubicin Resistant Breast Cancer

SignificanceEmploying mechanistically distinct therapies like Photodynamic Therapy (PDT) can be a promising strategy to overcome the multidrug- resistance in breast cancer resulting in the killing of chemotherapeutic drug-resistant cancer cells. ApproachIn this study, Pheophorbide a (Pheo-a), is being used as a potential photosensitizer to induce cell death in Doxorubicin (DOX) resistant breast cancer MCF-7 cells. Efficacy of Pheo-a mediated PDT against MCF-7/DOX cancer cells was evaluated by MTT proliferation assay and analyzing cellular morphological changes, live dead cell death assay. ResultsAll experiments were performed thrice in triplicate, and the results obtained were analysed for statistical significance. Results showed that the Pheo-a can induce photodynamic cell death in resistant cancer cells treated with a 2.5 μM concentration of Pheo-a followed by irradiation with a 660 nm red laser at a light dose of 10 J/cm2. ConclusionsThus, the results from this study suggest the utilization of chlorophyll-based PSs as a rational therapeutic intervention for the eradication of chemo drug-resistance in cancers.

Effectiveness of Pheophorbide-a mediated Photodynamic Therapy against Doxorubicin Resistant Breast Cancer

SignificanceEmploying mechanistically distinct therapies like Photodynamic Therapy (PDT) can be a promising strategy to overcome the multidrug- resistance in breast cancer resulting in the killing of chemotherapeutic drug-resistant cancer cells. ApproachIn this study, Pheophorbide a (Pheo-a), is being used as a potential photosensitizer to induce cell death in Doxorubicin (DOX) resistant breast cancer MCF-7 cells. Efficacy of Pheo-a mediated PDT against MCF-7/DOX cancer cells was evaluated by MTT proliferation assay and analyzing cellular morphological changes, live dead cell death assay. ResultsAll experiments were performed thrice in triplicate, and the results obtained were analysed for statistical significance. Results showed that the Pheo-a can induce photodynamic cell death in resistant cancer cells treated with a 2.5 μM concentration of Pheo-a followed by irradiation with a 660 nm red laser at a light dose of 10 J/cm2. ConclusionsThus, the results from this study suggest the utilization of chlorophyll-based PSs as a rational therapeutic intervention for the eradication of chemo drug-resistance in cancers.

Pediococcus acidilactici isolated from traditional cheese as a potential probiotic with cytotoxic activity against doxorubicin-resistant MCF-7 cells.

The considerable flexibility of cancerous cells to escape from chemical and biological drugs makes it clear that much is to be done to control and eliminate such cells. Probiotic bacteria, in this regard, have shown promising performance. In this study, we isolated and characterized lactic acid bacteria from traditional cheese. Then we evaluated their activity against doxorubicin-resistant MCF-7 cells (MCF-7/DOX) through MTT assay, Annexin V/PI protocol, real-time PCR, and western blotting. Among the isolates, one strain with more than 97% similarity with Pediococcus acidilactici showed considerable probiotics properties. Low pH, high bile salts, and NaCl could not significantly affect this strain while it was susceptible to antibiotics. Also, it had a potent antibacterial activity. Besides, the cell-free supernatant of this strain (CFS) significantly reduced the viability of MCF-7 and MCF-7/DOX cancerous cells (to about 10% and 25%, respectively), while it was safe for normal cells. Also, we found that CFS could regulate the Bax/Bcl-2 at mRNA and protein levels to induce apoptosis in drug-resistant cells. We determined 75% early apoptosis, 10% late apoptosis, and 15% necrosis in the cells treated with the CFS. These findings can accelerate the development of probiotics as promising alternative treatments to overcome drug-resistant cancers.

Docosahexaenoic Acid, a Key Compound for Enhancing Sensitization to Drug in Doxorubicin-Resistant MCF-7 Cell Line.

Drug resistance is a well-known and significant obstacle in the battle against cancer, rendering chemotherapy treatments often ineffective. To improve the effectiveness of chemotherapy, researchers are exploring the use of natural molecules that can enhance its ability to kill cancer cells and limit their spread. Docosahexaenoic acid (DHA), a lipid found in marine fish, has been shown to enhance the cytotoxicity of various anti-cancer drugs in vitro and in vivo. While the combined use of chemotherapeutic drugs with DHA demonstrated promising preliminary results in clinical trials, there is still a significant amount of information to be discovered regarding the precise mechanism of action of DHA. As the biological pathways involved in the chemosensitization of already chemoresistant MCF-7 cells are still not entirely unraveled, in this study, we aimed to investigate whether DHA co-treatment could enhance the ability of the chemotherapy drug doxorubicin to inhibit the growth and invasion of MCF-7 breast cancer cells (MCF-7/Dox) that had become resistant to the drug. Upon treating MCF-7/Dox cells with DHA or DHA-doxorubicin, it was observed that the DHA-doxorubicin combination effectively enhanced cancer cell death by impeding in vitro propagation and invasive ability. In addition, it led to an increase in doxorubicin accumulation and triggered apoptosis by arresting the cell cycle at the G2/M phase. Other observed effects included a decrease in the multi-drug resistance (MDR) carrier P-glycoprotein (P-gp) and TG2, a tumor survival factor. Augmented quantities of molecules promoting apoptosis such as Bak1 and caspase-3 and enhanced lipid peroxidation were also detected. Our findings in the cell model suggest that DHA can be further investigated as a natural compound to be used alongside doxorubicin in the treatment of breast cancer that is unresponsive to chemotherapy.

High Levels of Hyaluronic Acid Synthase-2 Mediate NRF2-Driven Chemoresistance in Breast Cancer Cells.

Hyaluronic acid (HA), a ligand of CD44, accumulates in some types of tumors and is responsible for tumor progression. The nuclear factor erythroid 2-like 2 (NRF2) regulates cytoprotective genes and drug transporters, which promotes therapy resistance in tumors. Previously, we showed that high levels of CD44 are associated with NRF2 activation in cancer stem like-cells. Herein, we demonstrate that HA production was increased in doxorubicin-resistant breast cancer MCF7 cells (MCF7-DR) via the upregulation of HA synthase-2 (HAS2). HA incubation increased NRF2, aldo-keto reductase 1C1 (AKR1C1), and multidrug resistance gene 1 (MDR1) levels. Silencing of HAS2 or CD44 suppressed NRF2 signaling in MCF7-DR, which was accompanied by increased doxorubicin sensitivity. The treatment with a HAS2 inhibitor, 4-methylumbelliferone (4-MU), decreased NRF2, AKR1C1, and MDR1 levels in MCF7-DR. Subsequently, 4-MU treatment inhibited sphere formation and doxorubicin resistance in MCF7-DR. The Cancer Genome Atlas (TCGA) data analysis across 32 types of tumors indicates the amplification of HAS2 gene is a common genetic alteration and is negatively correlated with the overall survival rate. In addition, high HAS2 mRNA levels are associated with increased NRF2 signaling and poor clinical outcome in breast cancer patients. Collectively, these indicate that HAS2 elevation contributes to chemoresistance and sphere formation capacity of drug-resistant MCF7 cells by activating CD44/NRF2 signaling, suggesting a potential benefit of HAS2 inhibition.

Rhinacanthin-C enhances chemosensitivity of breast cancer cells via the downregulation of P-glycoprotein through inhibition of Akt/NF-kappa B signaling pathway

Introduction: High expression of P-glycoprotein (P-gp) has been linked to multidrug resistance (MDR) and chemotherapeutic failure. Previously, we demonstrated that rhinacanthin-C, a naphthoquinone from Rhinacanthus nasutus, was able to enhance the cytotoxicity of doxorubicin against breast cancer MCF-7 cells via direct P-gp inhibition. In this study, we looked at its effect on P-gp downregulation and the mechanism involved in the resistance of MCF-7 cells to doxorubicin. Methods: Doxorubicin-resistant MCF-7 (MCF-7/DOX) cells were exposed to rhinacanthin-C for 24-48 hours prior to the assessment of their chemosensitivity via MTT assay, P-gp activity via calcein-AM uptake assay, P-gp expression, and signaling via qRT-PCR and western blot analyses. Results: Pretreatment with 1 µM of rhinacanthin-C for 48 hours significantly enhanced cytotoxicity of doxorubicin, as well as camptothecin and etoposide, to MCF-7/DOX cells. In the rhinacanthin-C-treated cells, reduction of MDR1 mRNA and P-gp levels and increased intracellular calcein were observed. Moreover, phosphorylation of Akt, NF-ᴋB and IκB-α, along with YB-1 expression, significantly decreased after 24-hour treatment with rhinacanthin-C. In contrast, the naphthoquinone had no effect on expression levels of ERK1/2 and phosphorylated ERK1/2 under similar conditions. Conclusion: Rhinacanthin-C, at a non-cytotoxic concentration (1 µM), could downregulate P-gp expression in MCF-7/DOX cells via the inhibition of the Akt/NF-ᴋB signaling pathway and YB-1 expression. Long-term exposure to this natural naphthoquinone may increase the chemosensitivity of cancer cells with MDR phenotype.

Anticancer effects of asiatic acid against doxorubicin-resistant breast cancer cells via an AMPK-dependent pathway in vitro

Background and purposeAsiatic acid is one of the active compounds isolated from Centella asiatica and has been used to treat many diseases, including hypertension, pulmonary fibrosis, and cancer. It exhibits anticancer effects in many cancers, such as ovarian, lung and colon cancer; however, its anticancer effects in breast cancer and the underlying mechanism are not fully understood. Chemoresistance is often induced after the use of chemotherapy, and it is a challenging problem in cancer therapy. The effects of asiatic acid on chemoresistance in breast cancer have never been studied. Therefore, the aim of the present study was to examine the anticancer effects of asiatic acid in doxorubicin-resistant breast cancer MCF-7 cells. MethodsThe cells were incubated with asiatic acid at 0-160 μM for 2-24 h. Cell viability and cytotoxicity were evaluated by 3-[4, 5-dimethyl-2-thiazolyl]-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays. Florescent images were taken using a confocal microscope. P-gp function and apoptosis assays were performed using flow cytometry. Caspase activity was measured with the Caspase-Glo™ Assay System. The phosphorylation and expression of relevant proteins were assessed by western blots. Molecular docking was performed and scored by AutoDock. Cellular thermal shift assay (CETSA) was applied for experimental valuation. ResultsOur data demonstrated that asiatic acid induced cell death in multiple ways, including reactive oxygen species production, adenosine triphosphate (ATP) content reduction, and adaptive immunity balance via intrinsic apoptosis, AMP-activated protein kinase (AMPK), programmed death-ligand 1 (PD-L1), and indirect nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) transcriptional pathways, using experimental validation and in silico analysis. Moreover, asiatic acid also enhanced the sensitivity of doxorubicin-resistant MCF-7 cells to doxorubicin by improving P-glycoprotein (P-gp) function. ConclusionsThis study provides evidence that asiatic acid has strong anticancer effects to reverse multidrug resistance and could be developed as a promising adjuvant drug for the treatment of chemoresistant cancer.

Lysophosphatidate Promotes Sphingosine 1-Phosphate Metabolism and Signaling: Implications for Breast Cancer and Doxorubicin Resistance.

Lysophosphatidate (LPA) and sphingosine 1-phosphate (S1P) promote vasculogenesis, angiogenesis, and wound healing by activating a plethora of overlapping signaling pathways that stimulate mitogenesis, cell survival, and migration. As such, maladaptive signaling by LPA and S1P have major effects in increasing tumor progression and producing poor patient outcomes after chemotherapy and radiotherapy. Many signaling actions of S1P and LPA are not redundant; each are vital in normal physiology and their metabolisms differ. In the present work, we studied how LPA signaling impacts S1P metabolism and signaling in MDA-MB-231 and MCF-7 breast cancer cells. LPA increased sphingosine kinase-1 (SphK1) synthesis and rapidly activated cytosolic SphK1 through association with membranes. Blocking phospholipase D activity attenuated the LPA-induced activation of SphK1 and the synthesis of ABCC1 and ABCG2 transporters that secrete S1P from cells. This effect was magnified in doxorubicin-resistant MCF-7 cells. LPA also facilitated S1P signaling by increasing mRNA expression for S1P1 receptors. Doxorubicin-resistant MCF-7 cells had increased S1P2 and S1P3 receptor expression and show increased LPA-induced SphK1 activation, increased expression of ABCC1, ABCG2 and greater S1P secretion. Thus, LPA itself and LPA-induced S1P signaling counteract doxorubicin-induced death of MCF-7 cells. We conclude from the present and previous studies that LPA promotes S1P metabolism and signaling to coordinately increase tumor growth and metastasis and decrease the effectiveness of chemotherapy and radiotherapy for breast cancer treatment.

Decrease of ABCB1 protein expression and increase of G1 phase arrest induced by oleanolic acid in human multidrug-resistant cancer cells.

Oleanolic acid (OA) is a natural compound that can be found in a number of edible and medicinal plants and confers diverse biological actions. However, the direct target of OA in human tumor cells remains poorly understood, preventing its application in clinical and health settings. A previous study revealed that overexpression of caveolin-1 in human leukemia HL-60 cells can increase its sensitivity to OA. The present study aimed to investigate the effects of OA on the doxorubicin-resistant human breast cancer MCF-7 cell line (MCF-7/DOX), harringtonine-resistant human leukemia HL-60 cells (HL-60/HAR) and their corresponding parental cell lines. Western blotting was performed to measure protein expression levels, whilst Cell Counting Kit-8 (CCK-8) assays, cell cycle analysis (by flow cytometry) and apoptosis assays (with Annexin V/PI staining) were used to assess drug sensitivity. CCK-8 assay results suggested that MCF-7/DOX cells, which overexpress the caveolin-1 protein, have similar OA susceptibility to their parent line. In addition, sensitivity of MCF-7/DOX cells to OA was not augmented by knocking down caveolin-1 using RNA interference. HL-60/HAR cells exhibited a four-fold increased sensitivity to OA compared with that in their parental HL-60 cells according to CCK-8 assay. Both of the resistant cell lines exhibited higher numbers of cells at G1 phase arrest compared with those in their parent lines, as measured via flow cytometry. Treatment of both MCF-7 cell lines with 100 µM OA for 48 h induced apoptosis, with increased effects observed in resistant cells. However, no PARP-1 or caspase-3 cleavage was observed, with some positive Annexin V staining found after HL-60/HAR cells were treated with OA, suggesting that cell death occurred via non-classical apoptosis or through other cell death pathways. It was found that OA was not a substrate of ATP-binding cassette subfamily B member 1 (ABCB1) in drug-resistant cells, as indicated by the accumulation of rhodamine 123 assessed using flow cytometry. However, protein expression of ABCB1 in both of the resistant cell lines was significantly decreased after treatment with OA in a concentration-dependent manner. Collectively, these results suggest that OA could reduce ABCB1 protein expression and induce G1 phase arrest in multidrug-resistant cancer cells. These findings highlight the potential of OA for cancer therapy.

Abstract P6-20-09: Pharmacological inhibition of TFF3 enhances chemo-sensitivity and overcomes acquired resistance in breast cancer

Abstract Background Dose-dependent toxicity and acquired chemo-resistance are two major challenges in the use of doxorubicin in breast cancer treatment. Trefoil factor 3 (TFF3) is a secreted ligand that promotes breast cancer progression and predicts poor survival outcome of breast cancer patients. It has also been shown to confer resistance to anti-estrogens and trastuzumab in breast cancer. Here, the role of TFF3 in regulating the sensitivity and acquired resistance to doxorubicin in breast cancer was investigated. Methods MCF7, ZR-75-1 and BT474 breast cancer cell lines with siRNA-mediated depletion of TFF3, and doxorubicin-resistant MCF7 cells generated from the pulsatile exposure to doxorubicin, were used as in vitromodels. We have developed a novel non-toxic small molecule inhibitor of TFF3 (AMPC) that binds specifically to cysteine 57 residue of dimeric TFF3 and promotes its dissociation to monomers thereby, inhibiting its dimeric functions such as proliferation and apoptosis. Here, the effects of AMPC in enhancing doxorubicin sensitivity and overcoming acquired doxorubicin resistance in breast cancer cells were also explored. Results Consistent with siRNA-mediated depletion of TFF3, pharmacological inhibition of TFF3 by AMPC enhanced doxorubicin-mediated decrease in cell viability, foci formation and 3D growth of the breast cancer cells, suggesting that TFF3 inhibition increased the sensitivity of these cells to doxorubicin treatment. Notably, AMPC combined with doxorubicin in a synergistic manner, enabling doxorubicin dose reduction for the same inhibitory effect. Doxorubicin-induced AKT activation has been reported to antagonize the effects of doxorubicin and promote its resistance in breast cancer. Here, the inhibition of TFF3 by AMPC was shown to reduce AKT activation. Mechanistically, AMPC co-treatment suppressed doxorubicin-induced AKT activation thereby enhancing doxorubicin-induced apoptosis, with an overall up-regulation of pro-apoptotic and down-regulation of anti-apoptotic proteins, as compared to doxorubicin monotherapy. TFF3 also mediated the acquired doxorubicin resistance in MCF7 cells. Elevated expression of TFF3 was observed in the doxorubicin-resistant MCF7 cells as compared to the parental MCF7 cells, while the inhibition of TFF3 by AMPC completely abrogated the resistant phenotype of these cells as shown in the cell viability, foci formation and 3D growth assays. In concordance with the elevated levels of TFF3, doxorubicin-resistant MCF7 cells also exhibited increased activation of AKT with reduced susceptibility to doxorubicin-induced apoptosis as compared to the parental MCF7 cells. Consistently, this was reversed with AMPC co-treatment, which suppressed the elevated levels of activated AKT in the doxorubicin-resistant MCF7 cells, resulting in the re-sensitization of these resistant cells to doxorubicin-induced apoptosis. Similar to that in the parental cells, AMPC also exhibited a synergistic inhibitory effect with doxorubicin in the doxorubicin-resistant MCF7 cells. Conclusion The pharmacological inhibition of TFF3 with AMPC is a potential therapeutic approach to reduce the dose-dependent toxicity and to overcome the acquired resistance of doxorubicin in breast cancer. Citation Format: Poh HM, Chong QY, Chen RM, Pandey V, Salundi B, Kumar AP, Lee SC, Lobie PE. Pharmacological inhibition of TFF3 enhances chemo-sensitivity and overcomes acquired resistance in breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P6-20-09.

An ingenious non-spherical mesoporous silica nanoparticle cargo with curcumin induces mitochondria-mediated apoptosis in breast cancer (MCF-7) cells.

Curcumin delivery to cancer cells is challenging due to its hydrophobic nature, low bio distribution and low availability. Many nano vehicles suffer from low stability and toxicity, and hence the prerequisite of a non-toxic nano vehicle with effective drug delivery is still being delved. The present study investigates the delivery efficiency of curcumin with non-spherical mesoporous silica nanoparticles (MSNAs). Their mechanism of drug delivery and signalling proteins activated to induce apoptosis was further explored in MCF-7 cells. A non-spherical MSN was synthesised, functionalised with PEI (MSNAP) and analysed its intracellular behaviour. Our result indicates that MSNAP was non-toxic until 20 µg/mL and likely localizes in cytoplasmic vesicles. On contrast, well-known MCM-41P induced autophagosome formation, indicating cellular toxicity. Curcumin was loaded on MSNAP and its effectiveness in inducing cell death was studied in MCF-7 and in MCF-7R cells. Curcumin loading on MSNAP induces better cell death with 30 µM curcumin, better than unbounded curcumin. Western blot analysis suggest, curcumin induce apoptosis through the activation of caspase 9, 6, 12, PARP, CHOP and PTEN. The cell survival protein Akt1 was downregulated by curcumin with and without the nanostructure. Interestingly, cleaved caspase 9 was activated in higher amount in nano-conjugated curcumin compared to the free curcumin. But other ER resident protein like IRE1α, PERK and GRP78 were downregulated indicating curcumin disturbs ER homeostasis. Further, electron microscopic analysis reveled that nanocurcumin induced apoptosis by disrupting mitochondria and nucleus. Our results with doxorubicin resistant MCF-7 cell lines confirm nanodelivery of doxorubicin and curcumin sensitised cells effectively at lesser concentration. Further docking studies of curcumin indicate it interacts with the apoptotic proteins through hydrogen bonding formation and with higher binding energy.

Hypoxia-induced gene expression pattern in doxorubicin resistant MCF7 cells

Purpose: To investigate hypoxia-induced gene expression pattern in doxorubicin-resistant human breast cancer cells (MCF7). Methods: Human breast cancer cells (MCF7) were exposed to 60 episodes of 8 h hypoxia thrice a week for three months. Chemo-resistance to doxorubicin was assessed using 3-(4,5-dimethylthiazol-2- yl)-2, 5-diphenyl tetrazolium bromide (MTT) cell proliferation assay. Real-time quantitative polymerase chain reaction (qRT-PCR) assay was performed to assess gene expression pattern in doxorubicinresistant cells on exposure to hypoxia. Results: Hypoxia significantly increased the resistance of MCF7 cells to doxorubicin, with a maximum of 16.42-fold enhancement after 25 episodes of 8-h hypoxia, while the resistance thereafter significantly decreased with prolonged episodes of hypoxia (p < 0.05). Gene expression analysis revealed significant changes in 42 genes. The expressions of 10 of these genes were significantly upregulated, while those of 32 genes were significantly down-regulated (p < 0.05). Cytochrome P450 family 1, subfamily A, member1 (CYP1A1) was the most conspicuous upregulated gene (13.32-fold), while breast cancer gene 1 (BRCA1) was the most down-regulated (8.23-fold). Gene expression analysis after 60 episodes of 8-h hypoxia revealed the upregulation of CYP1A1 (5.77-fold). Similarly, 27 genes were significantly down-regulated, with BRCA2 as the most down-regulated gene (8.11-fold). Topoisomerase (DNA) II alpha (TOP2A) was the most down-regulated among genes involved in drug metabolism and resistance (6.37-fold), while cyclin-dependent kinase 2 (CDK2) was the most profoundly downregulated among genes involved in cell cycle regulation (3.56-fold). Conclusion: These results indicate that development of resistance to doxorubicin by MCF7 cells after short-term hypoxia results from the upregulation of genes responsible for the metabolism of doxorubicin and for shifting the cells to alternative pathway driven principally by EGF and ESR2. The observed down-regulation is an adaptation of the MCF7 cells to survive under long-term hypoxia.

Long Non-Coding RNA &lt;i&gt;ENST00000508435&lt;/i&gt; Promotes Breast Cancer Chemoresistance Through Pyroptosis Inhibition

Chemotherapeutic resistance is a significant issue in the management of breast cancer patients, and the involvement of pyroptosis, a type of inflammatory cell necrosis, in cancer has recently been proposed. Here we explored the role of lncRNA ENS0508435 in pyroptosis and tumor chemoresistance. Our results showed that the ENS0508435 expression was significantly upregulated in doxorubicin-resistant MCF-7 cells (MCF-7/DOX) and that ENS0508435 knockdown decreased the resistance of MCF-7/DOX cells to doxorubicin. Moreover, ENS0508435 overexpression could inhibit doxorubicin-induced pyroptosis by reducing the expression of NLRP3, ASC, caspase-1, and GSDMD. Further evaluation of the mechanism demonstrated that ENS0508435 directly bound to hnRNPU, causing it to recruit EZH2, which repressed TRAF6 gene expression. Thus, TRAF6 was unavailable to combine with MALT1 and caspase-1, which attenuated pyroptosis. Altogether, the results indicated that ENS0508435 promoted breast cancer chemoresistance and was involved in pyroptosis inhibition through the hnRNPU-EZH2-TARF6-MALT1-caspase 1 pathway.

Anti-cancer effects of Rhizoma Curcumae against doxorubicin-resistant breast cancer cells

BackgroundChemotherapy is a primary approach in cancer treatment after routine surgery. However, chemo-resistance tends to occur with chemotherapy in clinic, resulting in poor prognosis and recurrence. Nowadays, Chinese medicine may shed light on design of new therapeutic modes to overcome chemo-resistance. Although Rhizoma Curcumae possesses anti-cancer activities in various types of cancers, the effects and underlying mechanisms of its bioactive components against chemo-resistance are not clear. Therefore, the present study aims to explore the potential effects of Rhizoma Curcumae on doxorubicin-resistant breast cancer cells.MethodsThe expression and function of ABC transporters in doxorubicin-resistant MCF-7 breast cancer cells were measured by western blotting and flow cytometry. Cell viability was detected using MTT assay. The combination index was analyzed using the CalcuSyn program (Biosoft, Ferguson, MO), based on the Chou–Talalay method.ResultsIn our present study, P-gp was overexpressed at protein level in doxorubicin-resistant MCF-7 cell line, but short of MRP1 and BCRP1. Essential oil of Rhizoma Curcumae and the main bioactive components were assessed on doxorubicin-resistant MCF-7 cell line. We found that the essential oil and furanodiene both display powerful inhibitory effects on cell viability, but neither of these is the specific inhibitor of ABC transporters. Moreover, furanodiene fails to enhance the efficacy of doxorubicin to improve multidrug resistance.ConclusionOverall, our findings fill the gaps of the researches on chemo-resistance improvement of Rhizoma Curcumae and are also beneficial for Rhizoma Curcumae being developed as a promising natural product for cancer adjuvant therapy in the future.

Quercetin reversed MDR in breast cancer cells through down-regulating P-gp expression and eliminating cancer stem cells mediated by YB-1 nuclear translocation.

Overexpression of P-glycoprotein (P-gp) plays an important role in mediating multidrug resistance (MDR), resulting in chemotherapy failure of tumor patients and enhancement of cancer stem cell characteristics. By preparing doxorubicin (Dox) resistant human breast cancer MCF-7 cells, here, we wanted to evaluate the effects of quercetin (Que) on MDR reversal activity and investigate its possible mechanism. MCF-7 and MCF-7/dox cells were respectively treated by Dox, paclitaxel (Pac), or vincristine (Vcr) with or without Que intervention for 24hr. Cell viability, cell apoptosis, cell cycle, intracellular drug accumulation, the expression of P-gp and Y-box binding protein 1 (YB-1), and breast cancer stem cells (BCSCs) were then assessed. The results showed that Que significantly enhanced the antitumor activities of Dox, Pac, and Vcr in breast cancer cells. In addition, combined treatment of Dox, Pac, or Vcr with Que significantly downregulated P-gp expression and eliminated BCSCs. Furthermore, combined treatment of Dox, Pac, or Vcr with Que significantly inhibited nuclear translocation of YB-1. Thus, we speculated that Que reversed MDR in breast cancer cells through downregulating P-gp expression and eliminating cancer stem cells mediated by YB-1 nuclear translocation.

Dual function of programmed cell death 10 (PDCD10) in drug resistance

Drug resistance, a major challenge in cancer chemotherapy, is a result of several mechanistic alterations including resistance to apoptosis. Apoptosis is a well-controlled cell death mechanism which is regulated by several signaling pathways. Alterations in structure, function, and expression pattern of the proteins involved in the regulation of apoptosis have been linked to drug resistance. Programmed Cell Death 10 (PDCD10) protein is recently associated with the regulation of cell survival and apoptosis. However, the role of PDCD10 in drug resistance has not been clearly established. Here, we aimed to figure out the role of PDCD10 in resistance to anti-cancer agents in different cell lines. We found that PDCD10 expression was cell- and anti-cancer agent-specific; down-regulated in doxorubicin- and docetaxel-resistant MCF7 cells while up-regulated in doxorubicin-resistant HeLa cells. Down-regulation of PDCD10 expression by siRNA in parental MCF7 cells increased the resistance while it increased sensitivity in doxorubicin-resistant HeLa cells. Similarly, over-expression of PDCD10 in parental HeLa cells increased the resistance to doxorubicin while it re-sensitized doxorubicin-resistant MCF7 cells. Moreover, the alterations in PDCD10 expression led to changes in caspase 3/7 activity and the levels of apoptosis-related genes. Our results point out a possible dual role of PDCD10 in drug resistance for the first time in the literature and emphasize PDCD10 as a novel target for reversal of drug resistance in cancer.

Corrigendum: Furanodiene Induces Extrinsic and Intrinsic Apoptosis in Doxorubicin-Resistant MCF-7 Breast Cancer Cells via NF-κB-Independent Mechanism.

[This corrects the article on p. 648 in vol. 8, PMID: 28959205.].

A dual-targeting DNA tetrahedron nanocarrier for breast cancer cell imaging and drug delivery

To enhance efficacy of chemotherapy and achieve real-time imaging of cancer cells, it is crucial to develop nanocarriers with targeted drug delivery capacity and fluorescence property for cancer theranostics. Herein, a dual-targeting DNA tetrahedron nanocarrier (MUC1-Td-AS1411) was constructed for breast cancer cell imaging and targeted drug delivery. This nanocarrier consisted of three components: (i) DNA tetrahedron core for multivalent conjugation of function ligands and loading doxorubicin (Dox); (ii) activatable MUC1 aptamer probe (MUC1-probe), formed by the hybridization of MUC1 aptamer sequence with fluorophore extended from one vertex and complementary sequence with quencher, for targeting and imaging MUC1 protein on cytomembrane; (iii) AS1411 aptamer, which was hybridized to the overhang on three vertexes via prolonged sequence, for binding to nucleolin. Firstly, MUC1-probe of this nanocarrier targeted MUC1 protein of MUC1-positive cells, causing a conformational reorganization of MUC1 aptamer, releasing complementary sequence with quencher and leading to fluorescence recovery. Subsequently, after internalizing into cells, AS1411 aptamer moiety of nanocarrier bound to nucleolin selectively, then the whole nanocarrier targeted nucleus and released Dox into nucleus. MUC1-positive cells and MUC1-negative cells could be differentiated by means of fluorescence imaging. Versus free Dox, Dox-loaded MUC1-Td-AS1411 showed lower cytotoxicity to MUC1-negative HL-7702 cells (P < 0.01), approximately equal lethality to sensitive MCF-7 cells (P > 0.05) whereas more effective to doxorubicin-resistant MCF-7 cells (P < 0.01). Therefore, this nanocarrier could be used as a promising candidate for cancer theranostics.

Furanodiene Induces Extrinsic and Intrinsic Apoptosis in Doxorubicin-Resistant MCF-7 Breast Cancer Cells via NF-κB-Independent Mechanism.

Chemotherapy is used as a primary approach in cancer treatment after routine surgery. However, chemo-resistance tends to occur when chemotherapy is used clinically, resulting in poor prognosis and recurrence. Currently, Chinese medicine may provide insight into the design of new therapies to overcome chemo-resistance. Furanodiene, as a heat-sensitive sesquiterpene, is isolated from the essential oil of Rhizoma Curcumae. Even though mounting evidence claiming that furanodiene possesses anti-cancer activities in various types of cancers, the underlying mechanisms against chemo-resistant cancer are not fully clear. Our study found that furanodiene could display anti-cancer effects by inhibiting cell viability, inducing cell cytotoxicity, and suppressing cell proliferation in doxorubicin-resistant MCF-7 breast cancer cells. Furthermore, furanodiene preferentially causes apoptosis by interfering with intrinsic/extrinsic-dependent and NF-κB-independent pathways in doxorubicin-resistant MCF-7 cells. These observations also prompt that furanodiene may be developed as a promising natural product for multidrug-resistant cancer therapy in the future.

Activation of Akt characterizes estrogen receptor positive human breast cancers which respond to anthracyclines.

Anthracyclines are key components of human breast cancer chemotherapy. Here, we explored the role of Akt signaling in anthracycline resistance.The antitumor activity of doxorubicin and Akt inhibitor A-443654 alone or combined was examined in estrogen receptor (ER) positive and negative human breast cancer cell lines. Further, we examined mRNA changes induced by anthracyclines in locally advanced breast cancers biopsied before and after treatment in two clinical trials.Doxorubicin increased Akt phosphorylation in ER positive MCF7 and T47D cell lines, with no effect in ER negative MDA-MB231 breast cancer cells. A-443654 was significantly more cytotoxic in doxorubicin-resistant compared to doxorubicin-naïve MCF7. This difference was not observed in MDA-MB231. Among 24 patients, AKT1 gene expression increased 24 hrs after the initial epirubicin exposure in ER positive tumors responding to therapy (n=6), as compared to ER positive non-responders (n=7) or ER negative tumors (n=11). In contrast, AKT1 mRNA changes after 16 weeks of doxorubicin were unrelated to clinical response and ER status (n=30).In conclusion, rapid Akt activation was observed in ER positive breast cancers which responded to anthracyclines. Increased cytotoxicity of A-443654 in doxorubicin-resistant MCF7 cells indicates a possible role for Akt inhibitors in ER positive breast cancers where chemoresistance evolves.