- Volume 13 Issue 12
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RNAi-induced K-Ras Gene Silencing Suppresses Growth of EC9706 Cells and Enhances Chemotherapy Sensitivity of Esophageal Cancer
- Wang, Xin-Jie (Department of Integrated Traditional Chinese and Western Medicine, The First Affiliated Hospital of Zhengzhou University) ;
- Zheng, Yu-Ling (Department of Oncology, The First Affiliated Hospital of Zhengzhou University) ;
- Fan, Qing-Xia (Department of Oncology, The First Affiliated Hospital of Zhengzhou University) ;
- Zhang, Xu-Dong (Department of Oncology, The First Affiliated Hospital of Zhengzhou University)
- Published : 2012.12.31
To analyze the growth, proliferation, apoptosis, invasiveness and chemotherapy sensitivity of EC9706 cells after K-Ras gene silencing, an expression carrier pSilencer-siK-Ras was constructed, and the EC9706 cell line was transfected using a liposome technique. Six groups were established: Control, siRNA NC (transfected with empty vector pSilencer2.1); Ras siRNA (transfected with pSilencer-siK-Ras2); Paclitaxel; Paclitaxel + siRNA NC; and Ras siRNA + Paclitaxel. After the treatment, RT-PCR, Western blotting, MTT assay, flow cytometry and the Transwell technique were used to assess expression of K-Ras mRNA and protein in EC9706 cells, as well as cell growth, proliferation, apoptosis and invasiveness. The effect of Paclitaxel chemotherapy was also tested. pSilencer-siK-Ras2 effectively down-regulated expression of K-Ras mRNA and protein in EC9706 cells, growth being significantly inhibited. Flow cytometry indicated obvious apoptosis of cells in the experimental group, with arrest in the G1 phase; cell migration ability was also reduced. After pSilencer-siK-Ras2 transfection or the addition of Paclitaxel, EC9706 cells were suppressed to different extents; the suppressive effect was strengthened by combined treatment. The results suggested that RNAi-induced K-Ras gene silencing could enhance chemotherapy sensitivity of esophageal cancer.
- Akkiprik M, Celikel CA, Düsünceli F, et al (2008). Relationship between overexpression of Ras p21 oncoprotein and K-Ras codon 12 and 13 mutations in Trukish colorectal cancer patients. Turk J Gastroenterol, 19, 22-7.
- Bivona TG, Quatela SE, Bodemann BO, et al (2006). PKC regulates a farne-syl-electro-soatic switch on K-Ras that promotes its association with Bcl-X2 on mitochondria and induces apoptosis [J]. Molecular Cell, 21, 481. https://doi.org/10.1016/j.molcel.2006.01.012
- Chen Y, Lan Z, Hui Q, Li B, Yang Y (2008). Expression of RASSF1A and CyclinD1 in tissues with gastric mucosa lesion and their relationship. World Chin J Digestology, 16, 2604-9.
- Cong DG, Wang SF, Zhang TW (2006). mRNA expression of RASSF1A in esophageal squamous cell carcinoma and clinical significance thereof. Nat Med J China, 86, 1624-7.
- Leung RK, Whittaker PA (2005). RNA interference:from gene silencing to gene-specific therapeutics. Pharmacol Ther, 107, 222-39. https://doi.org/10.1016/j.pharmthera.2005.03.004
- Li J, Zhao Z, Zhou Q, Wang L (2009). Expression of Ras protein in esophageal cancer and precancerous lesions. Chin J Clin Oncol Rehab, 6, 11-13.
- Lo PH, Xie D, Chan KC, et al (2007). Reduced expression of RASSF1A in esophageal and naso-pharyngeal carcinomas significantly correlates with tumor stage. Cancer Lett, 257, 199-205. https://doi.org/10.1016/j.canlet.2007.07.018
- Pijnenborg JM, Dam-de Veen GC, Kisters N, et al (2007). RASSF1A methylation and K-Ras and B-rafmutations and recurrent endometrial cancer. Ann Oncol, 18, 491-7.
- Pu H, Chen X, Li X, et al (2011). Expression of hMLH1 and RASSF1A in esophageal cancer tissues and its relationship with prognosis. World Chin J Digestol, 19, 1347-52.
- Quinlan MP, Settleman J (2008). A KRas-specific Function in cancer Initiation. Chin J Cancer, 27, 673.
- Riely GJ, Marks J, Pao W (2009). K-RAS mutations in non-small cell lung cancer. Proc Am Thorac Soc, 6, 201-5. https://doi.org/10.1513/pats.200809-107LC
- Schimanski CC, Zimmermann T, Schmidtmann I, et al (2010). K-Ras mutation status correlates with the expression of VEGFR1, VEGFR2, and PDGFR alpha in colorectal cancer. Int J Colorectal Dis, 25, 181-6. https://doi.org/10.1007/s00384-009-0843-7
- Shen YM, Yang XC, Yang C, Shen JK (2008). Enhanced therapeutic effects for human pancreatic cancer by application K-Ras and IGF-IR antisense Oligodeoxynucleotids. World J Gastroentero, 14, 5176-85. https://doi.org/10.3748/wjg.14.5176
- Song MS, Chang JS, Song SJ, et al (2005). The centrosomal protein RAS association domain family protein 1A (RASSF1A)-binding protein 1 regulates mitotic progression by recruiting RASSF1A to spindle poles. J Biol Chem, 280, 3920-7. https://doi.org/10.1074/jbc.M409115200
- Stella G, Rojas FL, Barone C, et al (2009). KRAS and BRAF mutational status and response to cetuximab combination therapy in advanced gastric cancer (GC) patients [C], ASCO GICancers Symposium, Abstr34.
- Tanaka T, Ishiguro H, Kuwabara Y, et al (2010). Vascular endothelial growth factor C in esophageal cancer correlates with lymph node metastasis and poor patient prognosis. J Exp Clin Cancer Res, 29, 83. https://doi.org/10.1186/1756-9966-29-83
- Wang X, Zheng Y, Fan Q (2011). Ras pathway and the activated expression of cyclin D1 in esophageal squamous cell carcinoma. J Third Military Med University, 33, 2626-8.
- Wang YX, Gao L, Ji ZZ (2007). Inhibitaary effects of antisense oligonucleoti-de specific to K-Ras pointmutation on the target gene expression in uman pancreatic carcinoma cells [J] Chin Med J, 120, 1448.
- Yoo YA, Na AR, Lee MS, et al (2006). RASSF1A suppresses oncogenic H-Ras-induced c-Jun N-terminal kinase activation. Int J Oncol, 29, 1541-7.
- Zhang TW, Wang SF, Cong DG, et al (2007). Suppression of RASSF1A gene on human esophageal carcinoma cells: experiments in vitroR and in vivo. National Med J China, 87, 1214-6.
- Zhong YQ, Xia ZS, Fu YR, et al (2010). Knockdown of Htert by siRNA suppresses growth of Capan-2 human pancreatic cancer cell via the inhibition of expressions of Bcl-2 and COX-2. J Dig Dis, 11, 176-84. https://doi.org/10.1111/j.1751-2980.2010.00433.x
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