Antisense oligonucleotide mediated knockdown of HOXC13 affects cell growth and induces apoptosis in tumor cells and over expression of HOXC13 induces 3D-colony formation

Abstract

HOXC13 is a homeobox containing gene that plays crucial roles in hair development and origin of replication. Herein, we investigated the biochemical functions of HOXC13 and explored its potential roles in tumor cell viability. We have designed a phosphorothioate based antisense-oligonucleotide that specifically knockdown HOXC13 in cultured cells. Cell viability and cytotoxicity assays demonstrated that HOXC13 is essential for cell growth and viability. Antisense-mediated knockdown of HOXC13 affected the cell viability and induced apoptosis in cultured tumor cells. HOXC13 regulates the expression of cyclins and antisense-mediated knockdown of HOXC13 resulted in cell cycle arrest and apoptosis in colon cancer cells. Finally over expression of HOXC13 resulted in 3D-colony formation in soft-agar assay indicating its potential roles in cell proliferation and tumorigenesis.

Bisphenol-A induces expression of HOXC6, an estrogen-regulated homeobox-containing gene associated with breast cancer

“…The Flag-HOXC6-CMV4 construct was transfected into HEK293 cells using Lipofectamine-2000 (Invitrogen) and stably transfected cells were isolated using G418 selection procedure as described by us earlier (16, 17). In brief, 5 µg of the Flag-HOXC6 construct was incubated with 8 µL of lipofectamine transfection reagent for 30 min in 500 µL DMEM at room temperature and the mixture was added to HEK293 cells (at 80% confluency).…”

“…The supernatant containing the RNA was subjected to phenol-chloroform extraction followed by ethanol precipitation of RNA by incubating 1h at −80 °C (17). Reverse transcription (RT) reactions were performed in a total volume of 25 µL of cDNA containing 500 ng of RNA, 2.4 µM of oligo dT (Promega), 100 units of MMLV reverse transcriptase, 1X first strand buffer (Promega), 100 µM each of dATP, dGTP, dCTP and dTTP (Invitrogen), 1 mM DTT, and 20 units of RNaseOut (Invitrogen).…”

“…HOX gene expression specifies the identity of the body segments along the anterior-posterior axis and secondary axis in animals (7). Recent studies demonstrate that HOX gene expressions are disrupted in various cancers (8–17). Many HOX genes appear to play critical roles in tumor cell proliferation, metastasis, and angiogenesis and the level of expression of these genes is correlated with patient outcome (17–20).…”

“…Recent studies demonstrate that HOX gene expressions are disrupted in various cancers (8–17). Many HOX genes appear to play critical roles in tumor cell proliferation, metastasis, and angiogenesis and the level of expression of these genes is correlated with patient outcome (17–20). Thus, HOX genes are emerging as key players in variety of cancer and are potential targets for disease diagnosis and novel therapy.…”

Effective cytotoxic activity of OSW-1 on colon cancer by inducing apoptosis in vitro and in vivo

“…Too much growth and too little death can lead to the disruption of homeostasis that may result in cancer (42). Previous studies have suggested that apoptosis, a universal cellular process, is the mechanism of many anticancer drugs (43)(44)(45)(46). Several studies have reported that OSW-1 can induce calcium-dependent apoptosis by damaging the mitochondrial membrane and disrupting cellular homeostasis or caspase-8-dependent cleavage of Bcl-2 in leukemia and Chinese hamster ovary (CHO) cells in vitro, respectively (34,35).…”