An ultrasonic nanobubble-mediated PNP/fludarabine suicide gene system: A new approach for the treatment of hepatocellular carcinoma

Abstract

ObjectiveThe purpose of this study is to generate an ultrasonic nanobubble (NB)-mediated purine nucleoside phosphorylase (PNP)/fludarabine suicide gene system for the treatment of human hepatocellular carcinoma (HCC).MethodsNBs were prepared from a mixture the phospholipids 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dipalmitoyl-sn-glycero-3-phosphate (DPPA), perfluoropropane gas and other materials using the high shear dispersion method. NBs treated with ultrasound irradiation functioned as a gene-transfer system, and a self-constructed suicide gene expression plasmid, pcDNA3.1(+)/PNP, treated with fludarabine functioned as a therapeutic gene. This system was used to determine the cytotoxic effects of PNP/fludarabine on HepG2 cells and SMMC7721 cells.Results1. NBs with a small diameter (208–416 nm) and at a high concentration and fine homogeneity were prepared under the optimal method. 2. The pcDNA3.1(+)/PNP plasmid was efficiently transfected into HCC cells using ultrasonic NBs. 3. At 0.75μg/ml fludarabine, PNP/fludarabine showed marked cytotoxic effects toward HepG2 and SMMC7721 cells. PNP/fludarabine achieved the same effect against both SMMC7721 and HepG2 cells but at a lower concentration of fludarabine for the latter. 4. Bystander effects: a 10–20% decrease in the cell survival rate was observed when only 5–10% of transfected cells were PNP positive.ConclusionsNBs constitute a non-toxic, stable and effective gene-delivery platform. The PNP/fludarabine suicide gene system inhibited the growth of HCC cells, induced HCC cell apoptosis, and caused a notable bystander effect at a low fludarabine concentration. This study establishes an important new method for miniaturizing microbubbles and improving a new NB-mediated approach for gene therapy of HCC.

Nanobubble Mediated Gene Delivery in Conjunction With a Hand-Held Ultrasound Scanner

“…Relatively low-frequency US was irradiation to induce release of siRNA micelles in tumor tissue from the siRNA loaded NBs and then effectively delivered into cancer cells. Furthermore, several promising data have been reported on liver gene therapy by combining various types of NBs and US (Wu et al, 2016;Zhang et al, 2018). Nevertheless, our animal sonoporation study also revealed induction of gene transfer in the liver after intravenous injection of lipid-based NBs.…”

A Novel Signature Based on mTORC1 Pathway in Hepatocellular Carcinoma

“…In terms of CACYBP , it has been verified that CACYBP can promote hepatocellular carcinoma progression in the absence of RNF41 -mediated degradation [ 22 ]. Moreover, a study [ 23 ] found that PNP /fludarabine suicide gene system induced HCC cell apoptosis and inhibited the growth of HCC cells. Although we found no evidence supporting the correlation between SKAP2 and HCC, it has been verified that SKAP2 promotes podosome formation to facilitate tumor-associated macrophage infiltration and metastatic progression [ 24 ].…”

A Novel Signature Based On Mtorc1 Pathway In Hepatocellular Carcinoma

“…In terms of CACYBP, it have been verified that CACYBP can promote hepatocellular carcinoma progression in the absence of RNF41 mediated degradation [22]. Moreover, a study [23] found that PNP/fludarabine suicide gene system induced HCC cell apoptosis and inhibited the growth of HCC cells. Although we found no evidence supporting the correlation between SKAP2 and HCC, it has been verified that SKAP2 promotes podosome formation to facilitates tumor-associated macrophage infiltration and metastatic progression [24].…”