Abstract
BackgroundGastric cancer is highly aggressive disease. Despite advances in diagnosis and therapy, the prognosis is still poor. Various genetic and molecular alterations are found in gastric cancer that underlies the malignant transformation of gastric mucosa during the multistep process of gastric cancer pathogenesis. The detailed mechanism of the gastric cancer development remains uncertain. In present study we investigated the potential role of stathmin1 gene in gastric cancer tumorigenesis and examined the usefulness of RNA interference (RNAi) targeting stathmin1 as a form of gastric cancer treatment.MethodsA lentiviral vector encoding a short hairpin RNA (shRNA) targeted against stathmin1 was constructed and transfected into the packaging cells HEK 293 T and the viral supernatant was collected to transfect MKN-45 cells. The transwell chemotaxis assay and the CCK-8 assay were used to measure migration and proliferation of tumor cells, respectively. Quantitative real-time PCR and western blotting were used to detect the expression levels of stathmin1.ResultsLentivirus mediated RNAi effectively reduced stathmin1 expression in gastric cells. Significant decreases in stathmin1 mRNA and protein expression were detected in gastric cells carrying lentiviral stathmin-shRNA vector and also significantly inhibited the proliferation, migration in gastric cancer cells and tumorigenicity in Xenograft Animal Models.ConclusionsOur findings suggest that stathmin1 overexpression is common in gastric cancer and may play a role in its pathogenesis. Lentivirus mediated RNAi effectively reduced stathmin1 expression in gastric cells. In summary, shRNA targeting of stathmin1 can effectively inhibits human gastric cancer cell growth in vivo and may be a potential therapeutic strategy for gastric cancer.
Highlights
We evaluated the state of stathmin1 protein expression in MKN-45 cell line by Western blot analysis (Figure 2A)
To determines the efficacies of viral vectors, viral supernatants prepared from either non-silencing-short hairpin RNA (shRNA) or stathmin1shRNA were added to gastric cancer cells
MKN-45 cells infected with non-silencing- shRNA and stathmin1 shRNA2 or untreated were injected into mice, stathmin shRNA2 transfected grew rapidly as compared to negative control (p < 0.05) (Figure 4A, B). These results demonstrate that in vivo tumor growth was inhibited by shRNA-mediated knockdown of stathmin1 expression in MKN-45 cell lines
Summary
Malignant transformation of gastric cells is the consequence of a multistep process involving different genetic and epigenetic changes in numerous genes in combination with host genetic background and environmental factors [6]. Stathmin is one of the microtubule-regulating proteins that play a critical role in the assembly and disassembly of the mitotic spindle [8,9,10,11]. It is highly expressed in a wide variety of human cancers, including leukemia, breast, prostate, and lung cancer, and provides an attractive target for cancer therapy [12,13,14,15]. Our study on esophageal squamous cell cancer revealed that stathmin is a predictor of survival in stage IIA esophageal squamous cell carcinoma after surgery [16]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
