Inhibitory Effect of Ursolic Acid on Proliferation and Migration of Renal Carcinoma Cells and Its Mechanism

Abstract

Background. Renal carcinoma is one of the most common malignant tumors in the urinary system. Autophagy can be both activated and inhibited in renal carcinoma, and it plays a double-edged role in the development of renal carcinoma. In the early stage of cancer, autophagy can suppress tumors. In the late stage, autophagy contributes to the survival of tumor cells in an unfavorable environment, and some autophagy-related proteins P62, LC3B, and beclin-1 have become indicators of the prognosis of patients with renal carcinoma. Aim. To demonstrate that ursolic acid activates autophagy in renal carcinoma 786-O cells by inhibiting the hedgehog signaling pathway. Methods. The effect of ursolic acid on the viability of 786-O cells was determined by the MTT method; the effect of ursolic acid on the proliferation and migration of 786-O cells was examined by crystalline violet staining and scratch assay, respectively. For the study of autophagy, we firstly screened the time points. Western blot assay was used to detect the expression level of autophagic protein P62 at different time points of ursolic acid on 786-O. Then, the Cell MeterTM Autophagy Assay Kit was used to detect the effect of different doses of ursolic acid on the autophagic fluorescence intensity of 786-O cells; the Western blot method was used to detect the effect of different doses of ursolic acid on the expression levels of LC3II and P62 proteins in 786-O cells. Further, AdPlus-mCherry-GFP-LC3B adenovirus transfection was used to detect the effect of ursolic acid on the autophagic flow of 786-O cells; ursolic acid was combined with the autophagy inhibitor chloroquine (CQ) to detect the expression level of autophagy protein LC3II by Western blot. In terms of mechanism, the effect of ursolic acid on hedgehog signaling pathway-related proteins in 786-O cells was detected by Western blot. Results. Ursolic acid inhibited the activity, proliferation, and migration of 786-O cells, enhanced the fluorescence intensity of autophagosomes in 786-O cells, increased the expression level of autophagy marker protein LC3II, and inhibited the expression level of P62 in a time and dose-dependent manner; ursolic acid activated the autophagic flow in 786-O cells, which showed that ursolic acid caused the accumulation of autophagic fluorescent spots and enhanced the fluorescence intensity of autophagosomes. Ursolic acid activated the autophagic flow in 786-O cells, as evidenced by the accumulation of autophagic fluorescent spots and enhanced fluorescence intensity of autophagosomes, and the combined use of the autophagy inhibitor CQ increased the expression level of LC3II compared to ursolic acid alone; ursolic acid decreased the expression levels of PTCH1, GLI1, SMO, SHH, and c-Myc and increased the expression level of Sufu in the hedgehog signaling pathway. Conclusion. Ursolic acid activates autophagy in renal carcinoma 786-O cells, probably by inhibiting hedgehog signaling pathway activity.

Therapeutic potential and novel formulations of ursolic acid and its derivatives: an updated review

“…In vivo and in vitro gastrointestinal cancer studies 40 have shown that UA influences the production of the protein caspase executioner (C‐3, C‐8 and C‐9) that is implicated in both internal and extrinsic routes of apoptosis. Furthermore, research on UA found that it increased the expression of death receptors in LNCap cells and the human colon cancer cell line HCT116 by triggering reactive oxygen species, c‐Jun N‐terminal kinase (JNK) and C/EBP homologous protein‐dependent pathways 16,41 . Tests on the human bladder cancer cell line T‐24 revealed that UA produced apoptosis via stressing the cells in the endoplasmic reticulum and inhibiting the Akt/NF‐κB signalling pathways, respectively.…”

“…The minimal inhibitory concentration (MIC) ranged from 32 to 526 g mL −1 . Anti‐inflammatory properties of natural extracts containing UA were reported to suppress the progression of several food‐related bacteria, yeasts, 40 and the protozoan ( Trypanosoma cruzi ) 16 closely connected with Leishmania infection 41 . It was recently discovered that UA possesses antifungal, antiviral, 42 antiparasitic and antibacterial properties 44 …”

“…Furthermore, research on UA found that it increased the expression of death receptors in LNCap cells and the human colon cancer cell line HCT116 by triggering reactive oxygen species, c-Jun N-terminal kinase (JNK) and C/EBP homologous protein-dependent pathways. 16,41 Tests on the human bladder cancer cell line T-24 revealed that UA produced apoptosis via stressing the cells in the endoplasmic reticulum and inhibiting the Akt/NF-κB signalling pathways, respectively.…”

Retracted: Inhibitory Effect of Ursolic Acid on Proliferation and Migration of Renal Carcinoma Cells and Its Mechanism

“…Computational Intelligence and Neuroscience has retracted the article titled "Inhibitory Efect of Ursolic Acid on Proliferation and Migration of Renal Carcinoma Cells and Its Mechanism" [1] due to concerns that the peer review process has been compromised. Following an investigation conducted by the Hindawi Research Integrity team [2], signifcant concerns were identifed with the peer reviewers assigned to this article; the investigation has concluded that the peer review process was compromised.…”

“…Computational Intelligence and Neuroscience has retracted the article titled “Inhibitory Effect of Ursolic Acid on Proliferation and Migration of Renal Carcinoma Cells and Its Mechanism” [ 1 ] due to concerns that the peer review process has been compromised.…”