ABSTRACT
Objective@#To investigate the effect and potential mechanism of HOXC8 on the radiosensitivity of non-small cell lung cancer cell line A549, aiming to provide novel ideas for clinical combined treatment.@*Methods@#The A549 cells with stable knockdown of HOXC8 were constructed by using lentivirus and validated by qPCR and Western blot. The radiosensitivity of A549 stable cell line was assessed by plate clone formation assay. The expression levels of TGF-β1 and the proteins in the downstream signal pathway after knockdown of HOXC8 were detected by Western blot.@*Results@#The A549 cells with stable knockdown of HOXC8 were successfully constructed. The viability and clonogenic capacity of A549 cells were significantly reduced after silencing HOXC8. Silencing HOXC8 also increased the sensitivity of A549 cells to radiotherapy and significantly inhibited the expression of TGF-β1 and p-Smad2/3 proteins in the downstream signaling pathway.@*Conclusion@#Silencing HOXC8 can increase the sensitivity of A549 cells to radiotherapy probably by inhibiting TGF-β1 signaling transduction. HOXC8 might play an important role in A549 cells.
ABSTRACT
@# Objective: To investigate the mechanism of lncRNA XIST (XIST) on modulating gastric cancer progression via regulating miR-337-3p/HOXC8 axis. Methods: A total of 58 cases of gastric cancer tissues and corresponding para-cancerous tissues resected from March 2013 to January 2018 in Department of General Surgery, Kailuan General Hospital of Tangshan City were collected for this study; in addition, human gastric cancer cell lines (AGS, MGC803, HGC27) and human gastric mucosal GES-1 cells were also collected. qPCR was used to detect the expressions of XIST and miR-337-3p in above mentioned gastric tissues and cell lines. XIST-knockdown vectors, miR-337-3p mimics, miR-337-3p inhibitor and HOXC8-overexpression vectors were transfected into AGS cells. The proliferation and invasion of AGS cells were detected by CCK-8 and Transwell experiments respectively, and the expression levels of HOXC8, E-cadherin, N-cadherin and vimentin were detected by WB. The targeting relationships between XIST, miR337-3p and HOXC8 were verified by dual-luciferase reporter gene assay. Results: XIST was up-regulated in gastric cancer tissues and cell lines (all P<0.01). XIST knockdown significantly inhibited proliferation, invasion and EMT of AGS cells (P<0.05 or P<0.01). Moreover, XIST directly interacted with miR-337-3p and down-regulated its expression, while HOXC8 was the target gene of miR-3373p. Furthermore, XIST knockdown suppressed proliferation, invasion and EMT ofAGS cells through up-regulating the inhibitory effect of miR-337-3p on HOXC8 (P<0.05 or P<0.01). Conclusion: XIST knockdown can suppress the proliferation, invasion and EMT of AGS cells, which may be related with down-regulation of HOXC8 by targeting miR-337-3p.
ABSTRACT
The protein encoded by the HOXC gene regulates multiple processes during embryonic development,including cell growth,differentiation,apoptosis,migration and angiogenesis.The HOXC gene is also involved in the occurrence and development of various human solid tumors,such as colorectal cancer,gastric cancer,liver cancer,breast cancer,endometrial cancer,etc.In addition,the HOXC gene is closely related to the clinical stage and distant metastasis of the tumor.Thus,the HOXC gene can be used as a biological predictor of tumor metastasis and prognosis.
ABSTRACT
As a member of homeobox gene family,HOXC is expressed in many organs and can regulate gene expression,cell differentiation and morphogenesis.Abnormality of its function is closely related to the prognosis of leukemia,breast cancer,renal cell carcinoma,prostate cancer and so on.
ABSTRACT
In order to examine whether the Hoxc8 protein can deliver nucleic acid into mammalian cells, we designed several Hoxc8-derived recombinant proteins to be synthesized as glutathione S-transferase (GST) fused forms in E.coli (GST-Hoxc8(1-242), containing a full length of Hoxc8; GST-Hoxc8(152-242), possessing a deletion of the acidic N-terminus of Hoxc8; GST-Hoxc8(149-208), which contained the homeodomain only). After labeling these proteins with Oregon 488, we examined their membrane transduction ability under the fluorescence microscope and verified that all three proteins showed similar transduction efficiency. The ability of the proteins to form in vitro protein-DNA complexes was analyzed on agarose gel; both GST-Hoxc8(1-242) and GST- Hoxc8(149-208) formed complexes. In contrast, the GST- Hoxc8(152-242) protein did not form a complex. The GST-Hoxc8(149-208) protein formed a complex with DNA at a mass ratio of 1 1 (DNA protein), and GST- Hoxc8(1-242) formed a complex at a mass ratio of 1 5. When the DNA (pDsRed1-C1) and protein complexes were added to culture media containing mammalian cells, the cells uptook the complexes, which was indicated by red fluorescence expression under the fluorescent microscope. These results indicate that recombinant Hoxc8 derivatives that harbor a homeodomain are able to traverse the mammalian cellular membrane. DNA that is bound to the recombinant derivatives can be carried across the membrane as well. This process could be applied in the development of a useful delivery vector for gene therapy in the future.