Effect of RNAi-mediated silencing of Livin gene on biological properties of colon cancer cell line LoVo

Abstract

ABSTRACT. This study aimed to investigate the effect of RNAimediated silencing of the Livin gene on biological properties of the colon cancer cell line LoVo. Interference vectors pSilencer4.1-Ll and pSilencer4.1-L2 targeting the Livin gene were constructed and transfected into LoVo cells. The expression of the Livin gene was determined by RT-PCR and Western blotting. The apoptosis, cell cycle, colony formation, proliferation of LoVo cells, as well as their sensitivity to cisplatin, were detected by flow cytometry, colony formation assay and MTT. Livin mRNA and protein expression in LoVo cells could be effectively silenced by pSilencer4.1-Ll but not pSilencer4.1-L2. In the pSilencer4.1-Ll transfection group, the apoptosis rate of LoVo cells was significantly higher than in the control group (24.2 ± 3.2 vs 8.1 ± 1.4%, P < 0.01), and after 72 h, cell proliferation was clearly decreased (about 70% inhibition). Compared with the control group, the colony formation rate in pSilencer4.1-Ll transfection group was obviously decreased (15 RNAi-mediated Livin gene silencing in LoVo cells ± 4.6 vs 85 ± 5.8%, P < 0.01), with increased proportion of S phase cells (45.7 ± 4.9 vs 28.0 ± 3.0%, P < 0.01), decreased proportion of G1 phase cells (43.0 ± 5.2 vs 62.8 ± 5.1%, P < 0.01), and increased sensitivity to cisplatin (apoptosis rate increased from 43.4 ± 6.9 to 65.3 ± 6.2%, P < 0.01). pSilencer4.1-Ll can effectively silence Livin gene expression in LoVo colon cancer cells, inhibit cell proliferation and colony formation, induce apoptosis, and enhance sensitivity to cisplatin.

Livin enhances chemoresistance in head and neck squamous cell carcinoma

“…Therefore, in the present study, we investigated the role of Livin in determining susceptibility to commonly used chemotherapeutic drugs, such as docetaxel, cisplatin and 5-FU, in human HNSCC cell lines. Although several studies have described that Livin contribute to the resistance of various chemotherapeutic drugs including cisplatin in various cancers (20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34), the present study is the first to demonstrate the correlation between Livin and chemoresistance in human HNSCC.…”

“…Livin, a recently discovered IAP, is composed of a single BIR domain and a RING motif (9). Livin has been implicated in chemoresistance in various cancers (20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34). It has been reported to play a role in resistance to cisplatin in bladder cancer, renal carcinoma, gastic cancer, hepatocellular carcinoma, lung adenocarcinoma/ non-small cell carcinoma, osteosarcoma, colon cancer and ovarian carcinoma (20)(21)(22)(23)(24)(25)(26)(27)(28)(29)31).…”

Room‐Temperature Alkynylation of Phosphine Oxides with Copper Acetylides: Practical Synthesis of Alkynylphosphine Oxides (Eur. J. Org. Chem. 4/2016)

“…The choice of these six cell lines was based on their distinct sensitivity to pro‐apoptotic stimuli, knowing that resistance of cancer cells to pro‐apoptotic stimuli represents one of the major failure of cancer chemotherapy 10. While Hs683,11 LoVo12 and MCF‐713 cancer cells display actual sensitivity to pro‐apoptotic stimuli, the U373,11, 14 A54915 and SKMEL‐2816 cancer cell lines display various levels of resistance to pro‐apoptotic stimuli.…”

Research Progress on the Livin Gene and Osteosarcomas

“…The critical role inhibitor of apoptosis proteins (IAPs) play in regulating cellular apoptosis has become the focus of present anti-tumor research. So far, 8 members of IAPs have been discovered in human body including NAIP, XIAP (ILP-1/MIHA), cIAP-1 (HIAP-2/MIHB), Livin (ML-IAP/KIAP), Survivin (TIAP), cIAP-2 (HIAP-1/MI-HC), ILP-2 (Ts-IAP) and Apollon (Bruce) (Zou et al, 2014), in which Livin can be applied as a new target for malignant tumor treatment (including osteosarcoma) due to its specifically high expression in tumor tissues. (RING).…”