Oncolytic Adenovirus with SPAG9 shRNA Driven by DD3 Promoter Improved the Efficacy of Docetaxil for Prostate Cancer


. 2022 Apr 30;2022:7918067.


doi: 10.1155/2022/7918067.


eCollection 2022.

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Meng Lu et al.


J Oncol.


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Abstract

Prostate cancer (PCa) is a common malignant tumor of the male urinary system and ranks the second in the causes of tumor-related deaths. Differential display code 3 (DD3) is a noncoding gene that is specifically expressed in PCa. High expression of sperm-associated antigen 9 (SPAG9) is closely related to tumorigenesis of PCa, and SPAG9 is a therapeutic target for PCa. In this study, a new oncolytic adenovirus DD3-ZD55-SPAG9 was constructed by using DD3 promoter to enhance the efficacy and safety of adenovirus. The combined use of DD3-ZD55-SPAG9 and docetaxel showed that DD3-ZD55-SPAG9 significantly improved the anti-tumor efficacy of docetaxel in PCa both in vitro and in vivo. The mechanism was related to the induction of tumor cell apoptosis and the inhibition of tumor cell invasion. In conclusion, DD3-ZD55-SPAG9 combined with docetaxel is an effective strategy for PCa therapy.

Conflict of interest statement

The authors declare that they have no conflicts of interest.

Figures


Figure 1



Figure 1

The cytotoxicity of DD3-ZD55-SPAG9 on prostate cancer and normal prostate cells. (a) Different titers of DD3-ZD55-SPAG9 inhibited the proliferation of PC-3 cells. (b) Different concentration gradient of DTX inhibited the proliferation of PC-3 cells. (c) Different titers of DD3-ZD55-SPAG9 inhibited the proliferation of DU-145 cells. (d) Different concentration gradient of DTX inhibited the proliferation of DU-145 cells. (e) Different combinations of treatment inhibited the proliferation of PC-3 cells. (f) Different combinations of treatment inhibited the proliferation of DU-145 cells. (g) CCK-8 assay indicated the inhibition of WPMY-1 cell proliferation by different treatments. Data were expressed as mean ± SD (n = 3).


Figure 2



Figure 2

Combination treatment inhibited PCa cell migration and invasion. (a) The scratch wound healing assay of the migration of PC-3 cells (100x). (b) The scratch wound healing assay of the migration of DU-145 cells (100x). (c) Histogram of wound healing area (%) of different treatment groups of PC-3 cells. (d) Histogram of wound healing area (%) of different treatment groups of DU-145 cells. (e) Transwell assay of the invasion of PC-3 cells (200x). (f) Transwell assay of the invasion of DU-145 cells (200x). (g) Histogram of the number of invasion cells of different treatment groups of PC-3 cells. (h) Histogram of the number of invasion cells of different treatment groups of DU-145 cells. (i) Western blot analysis of E-cadherin, vimentin, and MMP-2 in PC-3 cells. (j) Western blot analysis of E-cadherin, vimentin, and MMP-2 in DU-145 cells. (k) Quantitative analysis of E-cadherin levels in PC-3 and DU-145 cells. (l) Quantitative analysis of vimentin levels in PC-3 and DU-145 cells. (m) Quantitative analysis of MMP-2 levels in PC-3 and DU-145 cells. (1) PBS, (2) ZD55-EGFP, (3) DD3-ZD55-SPAG9, (4) ZD55-SPAG9, (5) DTX, and (6) DD3-ZD55-SPAG9+DTX. Data were expressed as mean ± SD (n = 3). p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 compared to the DD3-ZD55-SPAG9+DTX group.


Figure 3



Figure 3

Combined treatment promoted PCa cell apoptosis. (a) Hoechst 33258 staining of PC-3 cells (200x). (b) Hoechst 33258 staining of DU-145 cells. (c) Histogram of apoptosis rate of different treatment groups of PC-3 and DU-145 cells. Data were expressed as mean ± SD (n = 3). p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 compared to the DD3-ZD55-SPAG9+DTX group.


Figure 4



Figure 4

Combination therapy inhibits xenograft tumor growth. (a) Pictures of nude mice and each groups of xenograft tumors. (b) Growth curve of xenograft tumors in each group. (c) Histogram of tumor volume of each group on 28th day. Data were expressed as mean ± SD (n = 5). p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 compared to the DD3-ZD55-SPAG9+DTX group. (d) HE staining of xenografts of different groups (400x).


Figure 5



Figure 5

Combination therapy promoted tumor cell apoptosis in vivo. (a)TUNEL staining of xenografts of different groups (400x). (b) Histogram of apoptosis rate of each group. Data were expressed as mean ± SD (n = 3). p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 compared to the DD3-ZD55-SPAG9+DTX group.


Figure 6



Figure 6

Combined therapy regulated EMT-related protein expression. (a) Immunohistochemistry staining of SPAG9, E-cadherin, vimentin, and MMP-2 in xenografts of different groups (400x). (b) Analysis of SPAG9 staining density for each group. (c) Analysis of E-cadherin, vimentin, and MMP-2 staining density for each group. p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 compared to the DD3-ZD55-SPAG9+DTX group.

References

    1. Siegel R. L., Miller K. D., Jemal A. Cancer statistics, 2019. CA: a Cancer Journal for Clinicians . 2019;69(1):7–34. doi: 10.3322/caac.21551.



      DOI



      PubMed

    1. Armstrong A. J. Updates in advanced prostate cancer 2018. Prostate Cancer and Prostatic Diseases . 2018;21(4):449–450. doi: 10.1038/s41391-018-0100-7.



      DOI



      PubMed

    1. Grozescu T., Popa F. Prostate cancer between prognosis and adequate/proper therapy. Journal of Medicine and Life . 2017;10(1):5–12.



      PMC



      PubMed

    1. Nguyen-Nielsen M., Borre M. Diagnostic and therapeutic strategies for prostate cancer. Seminars in Nuclear Medicine . 2016;46(6):484–490. doi: 10.1053/j.semnuclmed.2016.07.002.



      DOI



      PubMed

    1. Niu Y., Guo C., Wen S., et al. ADT with antiandrogens in prostate cancer induces adverse effect of increasing resistance, neuroendocrine differentiation and tumor metastasis. Cancer Letters . 2018;439:47–55. doi: 10.1016/j.canlet.2018.09.020.



      DOI



      PubMed

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