Co-expression of ING4 and P53 enhances hypopharyngeal cancer chemosensitivity to cisplatin in vivo

Abstract

Hypopharyngeal cancer is a distinct type of malignant head and neck tumor, which exhibits low sensitivity to anti-cancer drugs. The importance of developing methods for reducing chemotherapy resistance, and improving and enhancing prognosis has previously been emphasized and is considered a challenge for effective clinical treatment of hypopharyngeal cancer. The current study investigated the effects of co-expression of inhibitor of growth protein 4 (ING4) and P53, a tumor suppressor gene, on chemosensitivity to cisplatin in human hypopharyngeal cancer xenografts in vivo, and the potential molecular mechanisms involved. A tumor model was established by injecting athymic nude mice with FADU human hypopharyngeal cancer cells. Five days after intratumoral and peritumoral injections of an empty adenoviral vector (Ad), Ad-ING4-P53, cisplatin, or a combination of Ad-ING4-P53 and cisplatin (Ad-ING4-P53 + cisplatin) every other day for 5 days, the mice were euthanized and their tumors, livers, and kidneys were removed. The tumor weights were used to calculate the inhibition rate, and the expression levels of ING4 and P53 were detected by reverse transcription-polymerase chain reaction. Additionally, apoptotic cells were detected using terminal deoxynucleotidyl transferase dUTP nick end labeling, and immunohistochemistry determined the levels ING4, P53, B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) protein expression. The results demonstrated increased expression of ING4 and P53 in the Ad-ING4-P53 groups compared with PBS and Ad groups, indicating successful introduction of the genes into the tumor cells. Notably, the Ad-ING4-P53 + cisplatin group exhibited a higher inhibition rate compared with the four other groups. The results of immunohistochemistry analysis demonstrated that Bax expression was increased and Bcl-2 was decreased in the Ad-ING4-P53 + cisplatin group. This suggested that the enhanced cisplatin chemosensitivity with Ad-ING4-P53 gene therapy in hypopharyngeal cancer xenografts may be associated with apoptosis induction through upregulation of Bax expression and downregulation of Bcl-2. The results of the present study indicated that gene therapy combined with cisplatin treatment may be a promising treatment for human hypopharyngeal cancer.

Efficacy of combining ING4 and TRAIL genes in cancer-targeting gene virotherapy strategy: first evidence in preclinical hepatocellular carcinoma

“…In specific, the combination therapy induced more robust activation and infiltration of NK cells, induction of apoptosis, and inhibition of angiogenesis and vascularization than monotherapy in vivo ( Figures 5 and 6 ). These in vivo findings support the previously published considerations that concomitant delivery of ING4 gene, 19 , 20 , 25 and likewise TRAIL gene, 8 , 10 , 13 , 14 with another potential tumoricidal gene is more effective in gene-based cancer therapy to overcome cancer resistance, and also re-highlight the importance of OAd in mediating potential cancer-targeting dual gene virotherapy strategy and providing a more meaningful therapeutic maneuver against multi-genic and highly resistant cancers, such as HCC, particularly if the two transgenes are well chosen. 7 , 8 , 9 , 10 , 11 As combination of two viruses is a major challenge for translation relevance compared with use of single virus, a future study should examine a single oncolytic virus co-expressing both TRAIL and ING4 genes to examine whether expression of both genes by a single vector system can still elicit more potent anticancer effect than a control vector expressing single therapeutic gene.…”

“… 3 , 4 , 5 , 6 , 7 ING4 has been lately identified as a potential candidate in gene-based cancer therapy to halt tumor growth and angiogenesis and to facilitate antitumor apoptosis, particularly when combined with additional anticancer gene or drugs. 15 , 19 , 20 , 21 , 22 , 23 , 24 , 25 In despite of this, its beneficial role in OAd-based cancer gene virotherapy is not well investigated yet and remains to be determined. Likewise, to the best of our knowledge there is no previous attention has been given to the potentiality of combining TRAIL; a potent antitumor apoptotic ligand that its importance in cancer combination therapies has been strongly recommended, 8 , 10 , 14 with ING4 to improve their overall anticancer response and overcome cancer resistance.…”

“… 19 At that respect, delivery of ING4 gene via adenoviral vectors into HCC, 20 , 21 lung carcinoma, 22 colorectal cancer, 23 osteosarcoma, 24 hypopharngeal cancer 25 and other modalities human cancers, was found to not only inhibit tumor growth, angiogenesis and invasion, but also regulate cell proliferation and apoptosis and facilitate tumor cell sensitivity to other adjuvant anticancer agents. 19 , 20 , 21 , 22 , 23 , 24 , 25 Nevertheless, most of these previous trials were based on using the traditional replication-deficient adenoviral vector to deliver ING4 gene, and their data were collectively highlighted the importance of combining ING4 gene with additional antitumor gene or chemotherapeutic drugs to achieve more potent antitumor effects than ING4 gene-based monotherapy. 19 , 20 , 21 , 22 , 23 , 24 , 25 By far, the utility and usefulness of ING4 in gene virotherapy strategy is still poorly understood.…”

“… 19 , 20 , 21 , 22 , 23 , 24 , 25 Nevertheless, most of these previous trials were based on using the traditional replication-deficient adenoviral vector to deliver ING4 gene, and their data were collectively highlighted the importance of combining ING4 gene with additional antitumor gene or chemotherapeutic drugs to achieve more potent antitumor effects than ING4 gene-based monotherapy. 19 , 20 , 21 , 22 , 23 , 24 , 25 By far, the utility and usefulness of ING4 in gene virotherapy strategy is still poorly understood. In addition to that, as both ING4 and TRAIL genes have their own distinct antitumor mechanisms; this in turn may suppose their possible augmenting benefit in cancer gene combination therapy.…”

Ultrasound‐guided intertumoral injection of contrast agents combined with human p53 gene for the treatment of breast cancer

“…In addition, p53 gene‐targeted mutations can knock out the p53 gene in rat embryonic stem (ES) cell chimeras in the ES cell genome [17]. As a transcription factor, p53 is the first member of the p53 family, eliciting genome maintenance by regulating cell cycle arrest, DNA repair, senescence and apoptosis occurring in response to a broad range of cellular stresses [18,19]. It has been reported that the survival of tumor cells lacking functional p53 is increased due to decreased apoptosis in these cells [20].…”