CMBs carrying PTX and CRISPR/Cas9 targeting C‑erbB‑2 plasmids interfere with endometrial cancer cells

Abstract

Development of combination therapy to decrease side effects of chemotherapeutic drugs and increase their utilization rate in combination with gene editing is a key research topic in tumor treatment. The present study aimed to investigate the effect of cationic microbubbles (CMBs) carrying paclitaxel (PTX) and C-erbB-2 knockout plasmid on the endometrial cancer cell line HEC-1A and to determine how C-erbB-2 regulates the function of endometrial cancer cells. Cells were treated with CMB, PTX, PTX-CMBs, cationic plasmid-carrying or cationic PTX-carrying plasmid groups. After verifying the most effective combination of PTX-CMBs and plasmids, HEC-1A cells were transfected. Reverse transcription-quantitative (RT-q)PCR and western blotting were used to measure C-erbB-2 and protein expression. After verifying C-erbB-2 knockout, invasion, healing, clone formation and proliferation of HEC-1A cells were assessed. Simultaneously, expression levels of the genes for P21, P27 , mammalian target of rapamycin ( mTOR ), and Bcl-2 associated death promoter ( Bad ) were measured by RT-qPCR. Compared with the PTX group, CMBs significantly enhanced the absorption efficiency of PTX by HEC-1A cells. C-erbB-2 knockout had an inhibitory effect on the proliferation, migration and invasion of HEC-1A cells; cell proliferation and invasion of the group carrying PTX and plasmids simultaneously were significantly weakened. The C-erbB-2 -knockout group exhibited increased expression of P21 and P27 . Simultaneously loading PTX and plasmid may be novel combination therapy with great potential. C-erbB-2 may regulate the proliferation of HEC-1A cells by downregulating expression of P21 and P27 .

Effect of CMB Carrying PTX and CRISPR/Cas9 on Endometrial Cancer Naked Mouse Model

“…According to our previous study [12], the particle sizes of CMB and PTX-CMB were 1.213 μm and 1.970 μm, respectively. e surface potential of PTX-CMB and CMB was 20.41 mV and 36.70 mV, respectively.…”

“…Furthermore, virus-mediated drug delivery systems can lead to off-target accumulation of lysates, thus limiting the in vivo application of virus-mediated methods. As for nonvirusmediated physical methods, such as microinjection, electroporation, and microinjection, high labor costs and harsh experimental conditions are required [12]. is study describes an efficient delivery system using CMBs and ultrasonic waves.…”

“…. Construction and physical measurement of CMB-CRISPR/Cas9, CMB-PTX, and CMB-PTX-CRISPR/Cas9 was performed by our previous study [12].…”

“…e loading control was GADPH from Sigma (USA). Use horseradish peroxidase (HRP)-conjugated secondary antibody (Sigma, USA) and incubate with cells (1 h, 25 °C) [8,12]. Band densities were quantified using the Licor Odyssey Infrared Imaging System (Licor Bio-science, Nebraska, USA).…”

“…is leads to an increase in cell permeability and an increase in the drug delivery efficiency. e positively charged surface of CMBs is filled with cationic gas, and negatively charged plasmid DNA can effectively bind to it, thereby increasing the loading rate of the plasmid [12]. erefore, CMBs can be used as materials to deliver drugs or genes.…”