Baculovirus-Mediated miRNA Regulation to Suppress Hepatocellular Carcinoma Tumorigenicity and Metastasis

Abstract

MicroRNA 122 (miR-122) is a tumor suppressor for hepatocellular carcinoma (HCC) but is lowly expressed in HCC cells. MiR-151 is aberrantly overexpressed in HCC cells and promotes HCC metastasis yet its roles on HCC tumorigenicity are unknown. To combat HCC tumorigenicity/metastasis, we developed Sleeping Beauty (SB)-based hybrid baculovirus (BV) vectors that expressed (i) miR-122 precursors (pre-miR-122), (ii) miR-151 sponges, or (iii) pre-miR-122 and miR-151 sponges. Transduction of aggressive HCC cells (Mahlavu) with the pre-miR-122-expressing BV tremendously enhanced miR-122 levels for >6 weeks, suppressed the levels of downstream effectors (e.g., ADAM10 and Bcl-w), proliferation, anchorage-independent growth, motility and migration/invasion in vitro. Intratumoral injection of the pre-miR-122-expressing BV attenuated the HCC growth/metastasis. The miR-151 sponges-expressing BV diminished the miR-151 levels for 6 weeks, enhanced RhoGDIA expression, suppressed RhoGTPases, as well as motility and migration/invasion of Mahlavu cells. Intratumoral injection of the miR-151 sponge-expressing BV impeded not only HCC metastasis but also cell proliferation, MMP expression and tumor growth in vivo. The BV co-expressing pre-miR-122 and miR-151 sponges also simultaneously enhanced miR-122 expression and inhibited miR-151, and conferred antitumor/anti-metastasis effects albeit lack of synergism. These data implicate the potentials of the SB-based hybrid BV for persistently modulating miRNA and suppressing HCC tumorigenicity/metastasis.

MiR‐122 modification enhances the therapeutic efficacy of adipose tissue‐derived mesenchymal stem cells against liver fibrosis

“…Given that enhanced IGFR signalling is implicated in rapid proliferation of activated HSCs , 122‐Exo‐mediated down‐regulation of this gene may contribute to the inhibition of HSC proliferation by miR‐122‐modified AMSCs. A similar growth inhibitory effect of miR‐122 by regulation of several tumorigenesis‐related proteins, including ADAM10, TFDP2 and E2F1, was previously demonstrated in hepatoma cells .…”

Improved calvarial bone repair by hASCs engineered with Cre/loxP-based baculovirus conferring prolonged BMP-2 and MiR-148b co-expression

“…It was recently shown that delivery of anti‐miR‐138 oligonucleotide into BMSC sheets (Yan et al ., ) or nanoparticle‐assisted delivery of miR‐199a‐5p agomirs into human BMSCs (Chen et al ., 2015) enhanced osteogenesis in vitro . However, these studies only demonstrated ectopic bone formation (Yan et al ., ; Chen X et al ., ). Conversely, knocking down miR‐31 in rat ASCs (Deng et al ., ) or rat BMSCs (Deng et al ., ) by antisense RNA expression improved in vivo repair after implantation into critical‐size calvarial defects in rats, but healed only ≈ 35% (Deng et al ., ) and ≈ 60% (Deng et al ., ) of the defect area, respectively.…”

“…This step was repeated until the plasmid, pTA‐8 × miR‐148b, contained eight tandem copies of pre‐miR‐148b (8 × miR‐148b). The 8 × miR‐148b fragment was digested with SalI / Xho I and subcloned into pUC57‐SD‐SA (Chen CL et al ., ) in between the chimaeric intron sequence containing the splicing donor (SD) and splicing acceptor (SA) to yield pUC57–8 × miR‐148b. Next, the SD‐8 × miR‐148b‐SA fragment was PCR‐amplified from pUC57–8 × miR‐148b using primers P3 and P4 (see the Supplementary material online, Table S1) and inserted into the BamH I /Stu I site in pBacLCW (Sung et al ., ) to yield pBac‐L148b.…”

Stage‐dependent involvement of ADAM10 and its significance in epileptic seizures

“…57 Several miRNAs targeting ADAM10 have been reported in pathological conditions, such as tumours and AD. [66][67][68][69][70][71][72][73][74][75][76][77][78][79][80][81][82] However, the miRNAs involved in these pathological conditions nearly overlap; hence, the involvement of miRNAs in ADAM10 regulation is most likely disease specific, necessitating the development of disease-specific strategies for treating ADAM10-related diseases ( Figure 1; Table 1). miRNA/ADAM10 pathways have been primarily defined in tumours and AD but are undetermined in epilepsy.…”