Neutralizing FGF4 protein in conditioned medium of IL-21-silenced HCT116 cells restores the migratory activity of the colorectal cancer cells

Abstract

The interleukin-21 (IL-21) protein was found to be expressed at an elevated level in clinical samples of colorectal cancer patients without or with a parasitic infection that were collected from Sudan in our previous study. The IL-21 gene in HT29 and HCT116 cells was then correlated to cell proliferation and cell migration, as well as the cellular mechanisms associated with gene expressions in our present study. Our results demonstrated that silencing the IL-21 gene in HCT116 cells increased the cytotoxic level and fibroblast growth factor-4 (FGF4) mRNA expression in the cancer cells. Moreover, specific gene silencing reduced the migration of cancer cells compared to non-silenced cancer cells. These events were not observed in IL-21-silenced HT29 cells. Neutralizing FGF4 in conditioned medium of IL-21-silenced HCT116 cells further increased the cytotoxic level and restored the migratory activity of HCT116 cells in the culture compared to silencing the IL-21 gene alone in the cancer cells. Our results indicate the importance of both silencing the IL-21 gene and co-expression of the FGF4 protein in HCT116 cells, which pave the way for the discovery of important factors to be used as biomarkers for the design of drugs or cost-effective supplements to effectively treat the patients having infectious disease and HCT116 cells of colorectal cancer simultaneously in the future.

IFN‑γ and IL‑18 in conditioned media of parasite‑infected host and IL‑21‑silenced colorectal cancer cells

“…3B ). As the cells transfected with scrambled siRNA did not exhibit any significant differences compared with non-transfected cells in our previous study ( 7 , 10 ), non-transfected cells were used as control. Evaluation of the mRNA expression level of the cell proliferation markers Ki67 and PCNA using RT-qPCR demonstrated that the mRNA expression levels of Ki67 and PCNA were decreased in the IL-21-silenced HCT116 cells compared with those in the controls.…”

“…This phenomenon may also explain the relationship between parasitic infection and colorectal cancer. IL-21 was the focus of the present study, as a high level of IL-21 expression has been detected in colorectal cancer cells, such as HT29 and HCT116( 7 ). In our previous study, a high level of IL-21 was also detected in the serum samples of patients diagnosed with colorectal cancer with a history of parasitic infections ( 24 ).…”

“…no. sc-29528; Santa Cruz Biotechnology, Inc.; as optimised in our previous study) with 100 µl siRNA transfection medium ( 7 ). Solution A was then directly added to Solution B and mixed gently by pipetting before mixing with 800 µl of siRNA transfection medium.…”

“…The current study also compared the levels of soluble factors in the conditioned media of IL-21-silenced colorectal cancer cells and the impact of the soluble factors on the mRNA expression level of cell proliferation markers in both the host cells post parasitic infection and colorectal cancer cells post silencing of the IL-21 gene. The IL-21 gene was silenced in a colorectal cancer cell line, as it has been indicated to reduce the proliferation of the cells, and it may also be used to study the molecular mechanisms of host-parasite interactions that cause chronic diseases with respect to colorectal cancer ( 7 ). The present study provided useful information on the fundamental molecular mechanisms of the host-parasite interactions that may aid in early diagnosis, novel prescription drugs and cost-effective strategies for the treatment of infectious disease-associated colorectal cancer in the future.…”

Serum Cytokine Analysis in Patients with Hepatocellular Carcinoma

“…Fibroblast growth factor 4 [FGF4] is a member of the FGF superfamily, which is involved in numerous stages of embryonic development and enhances multiple oncogenic activities, including tumor proliferation, invasion/migration, and colony initiation abilities [63,64]. Although there are some reports on the involvement of FGF4 in the processes of cancers, including lung adenocarcinoma, colorectal cancer, breast cancer, and ovarian cancer [65][66][67][68], little is known about the involvement of FGF4 in HCC. In this study, we found, for the first time, that FGF4 was highly expressed in HCC, suggesting a role as a novel serum biomarker for HCC.…”