Novel Tubeimoside I liposomal drug delivery system in combination with gemcitabine for the treatment of pancreatic cancer

Abstract

Aim: To evaluate the anti-pancreatic cancer effect of novel Tubeimoside I multifunctional liposomes combined with gemcitabine.Methods: Liposomes were prepared through the thin film hydration method, with evaluations conducted on parameters including encapsulation efficiency (EE%), particle size, polydispersity index (PDI), zeta potential (ZP), storage stability, and release over a 7-day period. The cellular uptake rate, therapeutic efficacy in vitro and in vivo and the role of immune microenvironment modulation were evaluated.Results: The novel Tubeimoside I multifunctional liposomal exhibited good stability, significant anti-cancer activity, and immune microenvironment remodeling effects. Furthermore, it showed a safety profile.Conclusion: This study underscores the potential of Novel Tubeimoside I multifunctional liposomal as a promising treatment option for pancreatic cancer.

Keywords: F18-FDG micro-PET/CT imaging; gemcitabine; liposomes; pancreatic cancer; tubular alfalfa glycosides I; tumor microenvironment.

Figure 1.

The strategic creation of a versatile Tub I-GEM-LPs for targeting tumors includes replacing cholesterol with Tubeimoside I in Tub I-GEM-LPs. This switch acts as both a barrier enhancer and chemotherapy supplement, offering the liposome a variety of advantages. These advantages encompass improved absorption by malignant cells, restructuring of the tumor environment and heightened ability to eradicate tumor cells. By incorporating Tubeimoside I into the liposome, we have created a versatile therapeutic delivery system with the potential to improve the effectiveness of cancer treatments.

Figure 2.

Characterization of Tubeimoside I liposomes. (A) Size distribution of Tub I-LPs and C-LPs was examined with nanoparticle sizer. (B) Transmission electron microscope images of Tubeimoside I liposomes (Tub I-LPs) and cholesterol liposome (C-LPs); scale bar = 100 nm. (C) Stability of two kinds liposome formulations kept in 4°C conditions monitored by nanoparticle sizer. (n = 3; mean ± SD).（D) Drug leakage profles of GEM loaded C-LPs and Tub I-LPs (n = 3; mean ± SD).

Figure 3.

The liposome internalization and cellular uptake in Panc 02 and MIA PaCa 02 cells was investigated. Qualitative analysis (A & B) and quantitative analysis (C & D) of the uptake of carboxyfluorescein Coumarin 6-loaded C-LPs and Tub I-LPs in Panc 02 and MIA PaCa 02 cells was conducted. The cells were treated with 500 ng/ml Coumarin 6-loaded liposomes and incubated at 37°C for 4 h (n = 3; mean ± SD).

Figure 4.

In vitro anticancer effect of Tub I-GEM-LPs. (A) The cytotoxic effect of various groups on Panc02 cells was evaluated. (n = 5, mean ± SD). (B) The study also examined the cytotoxicity of different groups on MIA PaCa cells (n = 5, mean ± SD). (C) Representative scatter plots of Annexin V/PI analysis were provided for Panc02 cells after drug treatments. (D) The percentage of cells with early, late, and complete apoptosis in Panc 02 cells (n = 3; mean ± SD). (E) Similar analysis was conducted for MIA PaCa 02 cells (n = 3; mean ± SD). *p < 0.05; **p < 0.01 and ***p < 0.001.

Figure 5.

(A) Tumor weights of excised tumors n = 5; mean ± SD). (B) Tumor volume of excised tumors (n = 5; mean ± SD). (C) Tumors were removed from C57 mice with Panc 02 tumors the day following the final injection (n = 5; mean ± SD). (D) micro-PET/CT visualization of tumor-bearing rats after treatment. (E) Mean SUV was calculated for both groups of hormonal rats. (n = 3; mean ± SD) (F) Hematoxylin and eosin stained sections of the heart, liver, lungs and kidneys were examined in representative samples (100 μm scale bar). Statistical significance was observed (p < 0.001).

Figure 6.

While employed as a chemotherapy adjuvant, Tub I assisted in reversing the immunosuppressive TME. (A) Flow cytometry analysis detected the presence of CD4⁺ and CD8⁺ T cells through the use of CD3 FITC, CD8 APC and CD4 PE markers in spleen tissues. (B) The histograms analysis of CD8⁺ T cells. (C) Flow cytometry gating analysis of CD11b+/Gr1 + MDSC cells in tumors. (D) The histograms analysis of CD11b+/Gr1 + MDSC cells in spleen tissues p < 0.05, **p < 0.01 and ***p < 0.001, (n = 5; mean ± SD).

Figure 7.

(A & B) Immunofluorescence was employed to uncover the quantity of MDSC and CD8⁺ cells in tumors of mice bearing the Panc 02 tumor. Scale bar: 100 μm. (C & D) Immunofluorescence detection of CD8⁺ and MDSC cell numbers histogram analysis. *p < 0.05; **p < 0.01 and ***p < 0.001, (n = 5; mean ± SD).

Figure 8.

(A) The Western blotting analysis determined the protein expression of Bax and Bcl-2, with β-actin serving as the loading control. (B & C) Band densities of the proteins were quantified using the Image J software. *p < 0.05; **p < 0.01; ***p < 0.001; mean ± S.D; n = 5.