Simultaneous gene silencing of <i>KRAS</i> and anti-apoptotic genes as a multitarget therapy

Werner, Kristin; Lademann, Franziska; Thepkaysone, May-Linn; Jahnke, Beatrix; Aust, Daniela E.; Kahlert, Christoph; Weber, Georg F.; Weitz, Jürgen; Grützmann, Robert; Pilarsky, Christian

doi:10.18632/oncotarget.6766

Cited by 12 publications

(8 citation statements)

References 35 publications

(30 reference statements)

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“…These results demonstrate that PANC-1 and MIA PaCa-2 cell growth is sustained on KRAS expression, and KRAS inhibition, but not TFEB inhibition, increased apoptosis. This is in good agreement with our published results of simultaneous gene silencing of KRAS and apoptotic genes (18). Both primary PDAC cell lines (PANC-1, MIA PaCa-2) express activating mutations of KRAS and inactivating mutations of the P53 gene (31).…”

Section: Discussionsupporting

confidence: 92%

“…The following sequences were targeted by siRNAs: KRAS 5'-GGCTATATTTACATGCTA CTA-3' [Eurofins MWG Operon, Ebersberg, Germany (18)], TFEB 5'-CACAACTTAATTGAAAGGAGA-3' (Qiagen), and KIF11 (Eg5) 5'-AACTGAAGACCTGAAGACAAT-3' as the positive control (19); and nonsense siRNA (Allstars, Qiagen) served as the negative control. All transfections were performed with a concentration of 30 nM per target gene and an incubation time of 48-72 h.…”

Section: Methodsmentioning

confidence: 99%

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Role of TFEB-driven autophagy regulation in pancreatic cancer treatment

Klein

Werner

Teske

et al. 2016

International Journal of Oncology

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Abstract. Autophagy pathways promote the growth of pancreatic ductal adenocarcinoma (PDAC), but the critical role is yet to be determined. Transcription factor EB (TFEB) centrally controls lysosomal and autophagy biogenesis. This study aimed to explore the role of TFEB for autophagy regulation in PDAC. We found that TFEB expression was significantly elevated in human PDAC samples (n=45), and localized to the cytoplasm and nucleus in 11 of 15 cases. In primary PDAC cell lines, TFEB nuclear expression was evident even under basal conditions, and further nuclear enrichment was achieved by starvation. Transient RNA interference reduced TFEB expression to 11-23%, but starvation-induced accumulation of the lipidated, autophagosome-associated LC3-II and the autophago-to-lysosome route was maintained after TFEB silencing. Likewise, gemcitabine treatment of the cancer cells augmented apoptosis and LC3-II as an indicator of autophagy, regardless of the TFEB expression levels. Moreover, the interplay of oncogenic KRAS with TFEB and autophagy was investigated. KRAS silencing caused PDAC cell apoptosis and a reciprocal increase in TFEB expression. This inverse correlation could be confirmed in published data sets of genetically engineered mouse models and human PDAC samples using the the Pubmed GEO and BioPortal databases, and was independent of KRAS mutation status. In conclusion, the central autophagy regulator TFEB is expressed and active in PDAC, but autophagy is sustained after TFEB knockdown, suggesting alternative bypass signaling. TFEB is dispensable for gemcitabine-induced cell death, but inversely correlated with KRAS expression.

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Section: Discussionsupporting

confidence: 92%

Section: Methodsmentioning

confidence: 99%

Role of TFEB-driven autophagy regulation in pancreatic cancer treatment

Klein

Werner

Teske

et al. 2016

International Journal of Oncology

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“…Thus far, most of the clinical trials in gene therapy have been targeting cancer treatment (64.4% of all gene therapy trials) [10, 11] with remarkable efficacies noted both in vitro and in vivo [12–16]. These strategies typically include tumor cell apoptosis induction, tumor suppressor gene reintroduction, immunomodulation, oncogene inactivation and sensitivity gene introduction [12, 17].…”

Section: Introductionmentioning

confidence: 99%

A folate receptor-targeted lipoplex delivering interleukin-15 gene for colon cancer immunotherapy

et al. 2016

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Interleukin-15 has been implicated as a promising cytokine for cancer immunotherapy, while folate receptor α (FRα) has been shown to be a potentially useful target for colon cancer therapy. Herein, we developed F-PLP/pIL15, a FRα-targeted lipoplex loading recombinant interleukin-15 plasmid (pIL15) and studied its antitumor effects in vivo using a CT26 colon cancer mouse model. Compared with control (normal saline) treatment, F-PLP/pIL15 significantly suppressed tumor growth in regard to tumor weight (P < 0.001) and reduced tumor nodule formation (P < 0.001). Moreover, when compared to other lipoplex-treated mice, F-PLP/pIL15-treated mice showed higher levels of IL15 secreted in the serum (P < 0.001) and ascites (P < 0.01). These results suggested that the targeted delivery of IL15 gene might be associated with its in vivo antitumor effects, which include inducing tumor cell apoptosis, inhibiting tumor proliferation and promoting the activation of immune cells such as T cells and natural killer cells. Furthermore, hematoxylin and eosin staining of vital organs following F-PLP/pIL15 treatment showed no detectable toxicity, thus indicating that intraperitoneal administration may be a viable route of delivery. Overall, these results suggest that F-PLP/pIL15 may serve as a potential targeting preparation for colon cancer therapy.

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“…The KRAS is a key signaling pathway in pancreatic cancer. Typical alterations in the molecular signaling include mutations in KRAS in 95% of pancreatic cancer [41] , [51] . Consistent with our results, previous studies have also shown that ECT2 is an oncogene which is overexpressed in pancreatic tumor tissues [52] .…”

Section: Discussionmentioning

confidence: 99%

Identification of Prognostic Biomarkers by Combined mRNA and miRNA Expression Microarray Analysis in Pancreatic Cancer

Liu

Yang

Taher

et al. 2018

Translational Oncology

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Pancreatic cancer is the fourth leading cause for cancer-related death, and early diagnosis is one key to improve the survival rate of this disease. Molecular biomarkers are an important method for diagnostic use in pancreatic cancer. We used data from three mRNA microarray datasets and a microRNA dataset (GSE16515, GSE15471, GSE28735, and GSE41372) to identify potential key genes. Differentially expressed genes (DEGs) and microRNAs (DEMs) were identified. Functional, pathway enrichment, and protein-protein interaction analyses were performed on common DEGs across all datasets. The target genes of the DEMs were identified. DEMs targets that were also DEGs were further scrutinized using overall survival analysis. A total of 236 DEGs and 21 DEMs were identified. There were a total of four DEGs (ECT2, NR5A2, NRP2, and TGFBI), which were also predicted target genes of DEMs. Overall survival analysis showed that high expression levels of three of these genes (ECT2, NRP2, and TGFBI) were associated with poor overall survival for pancreatic cancer patients. The basic expression of DEGs in pancreas stood lower level in various organ tissues. The expression of ECT2 and NRP2 was higher in different pancreatic cancer cell lines than normal pancreas cell line. Knockout of ECT2 by Crispr Cas9 gene editing system decreased proliferation and migration ability in pancreatic cancer cell line MiaPaCa2. In conclusion, we think that data mining method can do well in biomarker screening, and ECT2 and NRP2 can play as potential biomarker or therapy target by Crispr Cas9 in pancreatic cancer.

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Simultaneous gene silencing of KRAS and anti-apoptotic genes as a multitarget therapy

Cited by 12 publications

References 35 publications

Role of TFEB-driven autophagy regulation in pancreatic cancer treatment

Role of TFEB-driven autophagy regulation in pancreatic cancer treatment

A folate receptor-targeted lipoplex delivering interleukin-15 gene for colon cancer immunotherapy

Identification of Prognostic Biomarkers by Combined mRNA and miRNA Expression Microarray Analysis in Pancreatic Cancer

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