Eexpression And Significance Of The Hedgehog Signaling Pathway Factors In Tongue Cancer Cell Line TCA

ObjectivesTo investigate the expression of Hedgehog (Hh) signaling pathway factors in tongue cancer cell line TCA, and to explore the effects of Hh signaling on TCA cell’s proliferation and migration.MethodsThe proliferation and migration was detacted by MTT assay and wound healing assay after treated TCA cells with Hh signaling pathway inhibitor Cyclopamine. And to analyzed the expression level of Hh signaling pathway factors mRNA and protein by RT-PCR and Western blot, respectively.ResultsMTT assay identified that Cyclopamine inhibited the proliferation of TCA cells was dependent on Cyclopamine’s concentration. Wound healing assay suggested the distance of Cyclopamine groups healing time was longer than control group. The TCA cells of treatment with Cyclopamine 24h, RT-PCR results showed that the expression of Ptch mRNA in the Cyclopamine groups was lower than the control group, respectively, and the expression in the 20μM Cyclopamine group and 10μM Cyclopamine group was lower than in the 5μM Cyclopamine group. The expression of Smo mRNA in the Cyclopamine groups was lower than the control group and the mRNA level was dependent on Cyclopamine’s concentration. In the TCA cells treatment with Cyclopamine 48h, showed that the expression of Ptch mRNA in the Cyclopamine groups was lower than in control group. The expression of Smo mRNA in the Cyclopamine groups was lower than control group and the mRNA level was dependent on Cyclopamine’s concentration. The expression of Glil mRNA in the Cyclopamine groups was higher than the control group. In the TCA cellstreatment with Cyclopamine 72h, showed that the expression of Ptch mRNA in the Cyclopamine groups was lower than in control group, and there was no significant difference between Cyclopamine each groups.The TCA cells treatment with Cyclopamine 24h, Western-Blot results showed that the expression of Ptch protein in the Cyclopamine groups was lower than in control group, the protein level was dependent on Cyclopamine’s concentration. The expression of Smo protein in the Cyclopamine groups was lower than the control group, and the expression in the 10μM Cyclopamine group was lower than in the 20μM Cyclopamine group. In the TCA cells treatment with Cyclopamine 48h, the expression of Ptch protein and Smo protein in the Cyclopamine groups was lower than in control group, and the expression in the 10μM Cyclopamine group was higher than in the other Cyclopamine groups. In the TCA cells treatment with Cyclopamine 72h, the expression of Smo protein in the Cyclopamine groups was lower than in control group and the protein level was dependent on Cyclopamine’s concentration.ConclusionCyclopamin was specifically inhibited the TCA cells migration and proliferation. The Cyclopamin inhibited the expression of Smo mRNA and protein. The Hh signaling might be involved in the carcinogenesis and progression of tongue cancer.