Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2013 Nov;34(33):8459-68.
doi: 10.1016/j.biomaterials.2013.07.050. Epub 2013 Aug 8.

Combinational delivery of c-myc siRNA and nucleoside analogs in a single, synthetic nanocarrier for targeted cancer therapy

Yuan Zhang  1 , Lei Peng, Russell J Mumper, Leaf Huang
Affiliations

Combinational delivery of c-myc siRNA and nucleoside analogs in a single, synthetic nanocarrier for targeted cancer therapy

Yuan Zhang et al. Biomaterials. 2013 Nov.

Abstract

The treatment of aggressive non-small-cell lung cancer (NSCLC) depends on the creation of new therapeutic regimens in clinical settings. In this study, we developed a Lipid/Calcium/Phosphate (LCP) nanoparticle that combines chemotherapy with gene therapy. By encapsulating a chemodrug, gemcitabine monophosphate (GMP), and siRNA specific to the undruggable cMyc oncogene (cMyc siRNA) into a single nano-sized vesicle and systemically administering them to nude mice, we achieved potent anti-tumor activity in both subcutaneous and orthotopic models of NSCLC. The improvements in therapeutic response over either cMyc siRNA or GMP therapy alone, were demonstrated by the ability to effectively induce the apoptosis of tumor cells and the significant reduction of proliferation of tumor cells. The combination therapy led to dramatic inhibition of tumor growth, with little in vivo toxicity. Additionally, the current studies demonstrated the possibility of incorporating both nucleic acid molecules and phosphorylated small molecule drugs into the inner core of a single nanoparticle formulation. Co-encapsulation of an oncogene-modulating siRNA and a chemotherapeutic agent will allow simultaneous interruption of diverse anti-cancer pathways, leading to increased therapeutic efficacy and reduced toxicities.

Keywords: Apoptosis; Calcium phosphate nanoparticle; Gemcitabine monophosphate; Proliferation; c-Myc siRNA.

PubMed Disclaimer

Figures

Figure 1
Figure 1
TEM pictures of (A) cMyc-LCPs, (B) GMP-LCPs and (C) (GMP+cMyc)-LCPs. Scale bar = 100 nm.
Figure 2
Figure 2
Drug-loaded LCPs induced potent cytotoxicity in vitro on (A) H460 and (B) A549 cells. The cell viabilities of (A) H460 and (B) A549 cells were measured using an MTT assay after 48 h of exposure to cMyc-LCP-AA, GMP-LCP-AA, (GMP+cMyc)-LCP-AA, (CMP+Con)-LCP-AA, free GMP, and a GMP-LCP-AA and cMyc-LCP-AA mixture (each at half dose). Data are mean ± S.D. (n=3).
Figure 3
Figure 3
Analysis of tumor proteins after systemic treatments. Mice bearing H460 tumors were given three daily IV injections and analysis of tumor proteins was prepared 24 h after the final injection using a western blot technique.
Figure 4
Figure 4
Apoptosis of tumor cells induced in vivo after the systemic administration of different LCPs in (A, B) H460 subcutaneous xenografts and (C, D) A549 orthotopic xenografts. The percentage (%) of apoptotic cells in (B) H460 and (D) A549 xenograft models: (B) *p<0.01, cMyc-LCP-AA vs. control, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. control, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. cMyc-LCP-AA; **p <0.001, GMP-LCP-AA vs. control, GMP-LCP-AA vs. cMyc- LCP-AA; ***p<0.0001, (GMP+cMyc)-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. cMyc-LCP-AA; (D) *p<0.0001, cMyc-LCP-AA vs. (GMP+cMyc)-LCP-AA, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA; **p<0.00001, cMyc-LCP-AA vs. control, GMP-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control. (n=5 per group)
Figure 4
Figure 4
Apoptosis of tumor cells induced in vivo after the systemic administration of different LCPs in (A, B) H460 subcutaneous xenografts and (C, D) A549 orthotopic xenografts. The percentage (%) of apoptotic cells in (B) H460 and (D) A549 xenograft models: (B) *p<0.01, cMyc-LCP-AA vs. control, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. control, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. cMyc-LCP-AA; **p <0.001, GMP-LCP-AA vs. control, GMP-LCP-AA vs. cMyc- LCP-AA; ***p<0.0001, (GMP+cMyc)-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. cMyc-LCP-AA; (D) *p<0.0001, cMyc-LCP-AA vs. (GMP+cMyc)-LCP-AA, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA; **p<0.00001, cMyc-LCP-AA vs. control, GMP-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control. (n=5 per group)
Figure 4
Figure 4
Apoptosis of tumor cells induced in vivo after the systemic administration of different LCPs in (A, B) H460 subcutaneous xenografts and (C, D) A549 orthotopic xenografts. The percentage (%) of apoptotic cells in (B) H460 and (D) A549 xenograft models: (B) *p<0.01, cMyc-LCP-AA vs. control, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. control, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. cMyc-LCP-AA; **p <0.001, GMP-LCP-AA vs. control, GMP-LCP-AA vs. cMyc- LCP-AA; ***p<0.0001, (GMP+cMyc)-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. cMyc-LCP-AA; (D) *p<0.0001, cMyc-LCP-AA vs. (GMP+cMyc)-LCP-AA, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA; **p<0.00001, cMyc-LCP-AA vs. control, GMP-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control. (n=5 per group)
Figure 4
Figure 4
Apoptosis of tumor cells induced in vivo after the systemic administration of different LCPs in (A, B) H460 subcutaneous xenografts and (C, D) A549 orthotopic xenografts. The percentage (%) of apoptotic cells in (B) H460 and (D) A549 xenograft models: (B) *p<0.01, cMyc-LCP-AA vs. control, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. control, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. cMyc-LCP-AA; **p <0.001, GMP-LCP-AA vs. control, GMP-LCP-AA vs. cMyc- LCP-AA; ***p<0.0001, (GMP+cMyc)-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. cMyc-LCP-AA; (D) *p<0.0001, cMyc-LCP-AA vs. (GMP+cMyc)-LCP-AA, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA; **p<0.00001, cMyc-LCP-AA vs. control, GMP-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control. (n=5 per group)
Figure 5
Figure 5
Proliferation of tumor cells in vivo after administration of different LCPs in (A, B) H460 subcutaneous xenografts and (C, D) A549 orthotopic xenografts. Twenty-four h after 3 daily IV injections, mice were sacrificed and tumors tissues were sectioned for PCNA immunohistochemistry. The percentage (%) of PCNA positive cells in (B) H460 and (D) A549 xenograft models was shown as follows: (B) *p<0.05, GMP-LCP-AA vs. control, GMP-LCP-AA vs. cMyc-LCP-AA, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA; **p <0.01, cMyc-LCP-AA vs. (GMP-LCP-AA)+(cMyc-LCPAA), (GMP+cMyc)-LCP-AA vs. (GMP-LCP-AA)+(cMyc-LCP-AA); ***p<0.001, cMyc-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control; (D) *p<0.01, cMyc-LCP-AA vs. (GMP+cMyc)-LCP-AA, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA; **p <0.0001, cMyc-LCP-AA vs. control, GMP-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control. (n=5 per group)
Figure 5
Figure 5
Proliferation of tumor cells in vivo after administration of different LCPs in (A, B) H460 subcutaneous xenografts and (C, D) A549 orthotopic xenografts. Twenty-four h after 3 daily IV injections, mice were sacrificed and tumors tissues were sectioned for PCNA immunohistochemistry. The percentage (%) of PCNA positive cells in (B) H460 and (D) A549 xenograft models was shown as follows: (B) *p<0.05, GMP-LCP-AA vs. control, GMP-LCP-AA vs. cMyc-LCP-AA, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA; **p <0.01, cMyc-LCP-AA vs. (GMP-LCP-AA)+(cMyc-LCPAA), (GMP+cMyc)-LCP-AA vs. (GMP-LCP-AA)+(cMyc-LCP-AA); ***p<0.001, cMyc-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control; (D) *p<0.01, cMyc-LCP-AA vs. (GMP+cMyc)-LCP-AA, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA; **p <0.0001, cMyc-LCP-AA vs. control, GMP-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control. (n=5 per group)
Figure 5
Figure 5
Proliferation of tumor cells in vivo after administration of different LCPs in (A, B) H460 subcutaneous xenografts and (C, D) A549 orthotopic xenografts. Twenty-four h after 3 daily IV injections, mice were sacrificed and tumors tissues were sectioned for PCNA immunohistochemistry. The percentage (%) of PCNA positive cells in (B) H460 and (D) A549 xenograft models was shown as follows: (B) *p<0.05, GMP-LCP-AA vs. control, GMP-LCP-AA vs. cMyc-LCP-AA, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA; **p <0.01, cMyc-LCP-AA vs. (GMP-LCP-AA)+(cMyc-LCPAA), (GMP+cMyc)-LCP-AA vs. (GMP-LCP-AA)+(cMyc-LCP-AA); ***p<0.001, cMyc-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control; (D) *p<0.01, cMyc-LCP-AA vs. (GMP+cMyc)-LCP-AA, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA; **p <0.0001, cMyc-LCP-AA vs. control, GMP-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control. (n=5 per group)
Figure 5
Figure 5
Proliferation of tumor cells in vivo after administration of different LCPs in (A, B) H460 subcutaneous xenografts and (C, D) A549 orthotopic xenografts. Twenty-four h after 3 daily IV injections, mice were sacrificed and tumors tissues were sectioned for PCNA immunohistochemistry. The percentage (%) of PCNA positive cells in (B) H460 and (D) A549 xenograft models was shown as follows: (B) *p<0.05, GMP-LCP-AA vs. control, GMP-LCP-AA vs. cMyc-LCP-AA, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA; **p <0.01, cMyc-LCP-AA vs. (GMP-LCP-AA)+(cMyc-LCPAA), (GMP+cMyc)-LCP-AA vs. (GMP-LCP-AA)+(cMyc-LCP-AA); ***p<0.001, cMyc-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control; (D) *p<0.01, cMyc-LCP-AA vs. (GMP+cMyc)-LCP-AA, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA; **p <0.0001, cMyc-LCP-AA vs. control, GMP-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control. (n=5 per group)
Figure 6
Figure 6
Inhibition of tumor growth in H460, subcutaneous xenografts of NSCLC. (A) Inhibition of tumor growth by different LCP formulations on H460-tumor bearing mice. cMyc-LCP-AA, GMP-LCP-AA, (GMP+cMyc)-LCP-AA, (CMP+Con)-LCP-AA and free GMP were administered intravenously every other day for total 4 injections. Tumor volumes were measured every other day. Data are mean ± S.D. (n=5) Statistics are as follows: *p<0.0005, GMPLCP- AA vs. control, cMyc-LCP-AA vs. control, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. control; **p <0.00001, (GMP+cMyc)-LCP-AA vs. control; ‡p<0.05, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA, cMyc-LCP-AA vs. (GMP+cMyc)-LCP-AA, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. (GMP+cMyc)-LCP-AA. (B) Weights of tumors collected from treated mice. Data are expressed as mean ± S.D. (n=5) Statistics are as follows: *p<0.005, cMyc-LCP-AA vs. control, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. control; **p <0.001, GMP-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control; ***p<0.01, cMyc-LCP-AA vs. (GMP+cMyc)-LCP-AA; ‡p<0.05, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. (GMP+cMyc)-LCP-AA. (C) The relative body weight changes over the treatment period of mice bearing H460 tumors. Data are mean ± S.D. (n=5)
Figure 6
Figure 6
Inhibition of tumor growth in H460, subcutaneous xenografts of NSCLC. (A) Inhibition of tumor growth by different LCP formulations on H460-tumor bearing mice. cMyc-LCP-AA, GMP-LCP-AA, (GMP+cMyc)-LCP-AA, (CMP+Con)-LCP-AA and free GMP were administered intravenously every other day for total 4 injections. Tumor volumes were measured every other day. Data are mean ± S.D. (n=5) Statistics are as follows: *p<0.0005, GMPLCP- AA vs. control, cMyc-LCP-AA vs. control, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. control; **p <0.00001, (GMP+cMyc)-LCP-AA vs. control; ‡p<0.05, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA, cMyc-LCP-AA vs. (GMP+cMyc)-LCP-AA, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. (GMP+cMyc)-LCP-AA. (B) Weights of tumors collected from treated mice. Data are expressed as mean ± S.D. (n=5) Statistics are as follows: *p<0.005, cMyc-LCP-AA vs. control, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. control; **p <0.001, GMP-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control; ***p<0.01, cMyc-LCP-AA vs. (GMP+cMyc)-LCP-AA; ‡p<0.05, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. (GMP+cMyc)-LCP-AA. (C) The relative body weight changes over the treatment period of mice bearing H460 tumors. Data are mean ± S.D. (n=5)
Figure 6
Figure 6
Inhibition of tumor growth in H460, subcutaneous xenografts of NSCLC. (A) Inhibition of tumor growth by different LCP formulations on H460-tumor bearing mice. cMyc-LCP-AA, GMP-LCP-AA, (GMP+cMyc)-LCP-AA, (CMP+Con)-LCP-AA and free GMP were administered intravenously every other day for total 4 injections. Tumor volumes were measured every other day. Data are mean ± S.D. (n=5) Statistics are as follows: *p<0.0005, GMPLCP- AA vs. control, cMyc-LCP-AA vs. control, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. control; **p <0.00001, (GMP+cMyc)-LCP-AA vs. control; ‡p<0.05, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA, cMyc-LCP-AA vs. (GMP+cMyc)-LCP-AA, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. (GMP+cMyc)-LCP-AA. (B) Weights of tumors collected from treated mice. Data are expressed as mean ± S.D. (n=5) Statistics are as follows: *p<0.005, cMyc-LCP-AA vs. control, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. control; **p <0.001, GMP-LCP-AA vs. control, (GMP+cMyc)-LCP-AA vs. control; ***p<0.01, cMyc-LCP-AA vs. (GMP+cMyc)-LCP-AA; ‡p<0.05, GMP-LCP-AA vs. (GMP+cMyc)-LCP-AA, (GMP-LCP-AA)+(cMyc-LCP-AA) vs. (GMP+cMyc)-LCP-AA. (C) The relative body weight changes over the treatment period of mice bearing H460 tumors. Data are mean ± S.D. (n=5)
See this image and copyright information in PMC

References

    1. de Nigris F, Balestrieri ML, Napoli C. Targeting c-Myc, Ras and IGF cascade to treat cancer and vascular disorders. Cell Cycle. 2006;5(15):1621–1628. - PubMed
    1. Dang CV. MYC on the path to cancer. Cell. 2012;149(1):22–35. - PMC - PubMed
    1. Toyoshima M, Howie HL, Imakura M, Walsh RM, Annis JE, Chang AN, et al. Functional genomics identifies therapeutic targets for MYC-driven cancer. Proc Natl Acad Sci U S A. 2012;109(24):9545–9550. - PMC - PubMed
    1. Zhang Y, Kim WY, Huang L. Systemic delivery of gemcitabine triphosphate via LCP nanoparticles for NSCLC and pancreatic cancer therapy. Biomaterials. 2013;34(13):3447–3458. - PMC - PubMed
    1. Oguri T, Achiwa H, Sato S, Bseeho Y, Takano Y, Miyazaki M, et al. The determinants of sensitivity and acquired resistance to gemcitabine differ in non-small cell lung cancer: a role of ABCC5 in gemcitabine sensitivity. Mol Cancer Ther. 2006;5(7):1800–1806. - PubMed

Publication types