Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene

Abstract

Objective. Tenacissoside H (TDH) is a Chinese medicine monomer extracted from Marsdenia tenacissima extract (MTE), which has been confirmed to have antitumor effects, but its mechanism is still unclear. The aim of this study was to investigate the effect and mechanism of TDH on human colon cancer LoVo cell proliferation and migration and explore the correlation of TDH treatment with the expression of GOLPH3 and cell signaling pathways in LoVo cells. Methods. LoVo cells were treated with TDH at 0.1, 1, 10, and 100 μg/mL for 24, 48, and 72 h. The proliferation rate of LoVo cells was evaluated by MTT assay. Recombinant plasmid p-CMV-2-GOLPH3 was constructed, and p-CMV-2-GOLPH3 and p-CMV-2 empty plasmids were transfected into LoVo cells by lipofection. Western blotting was used to detect the transfection efficiency and the expression of p-p70S6K, p70S6K, β-catenin, and GOLPH3. The apoptosis rate was analyzed with Annexin V-FITC/PI double-staining method, and cell migration assessed by transwell assay. Results. TDH inhibited the proliferation of LoVo cells in a concentration-dependent manner. The IC50 of TDH treatment in LoVo cells at 24, 48, and 72 h was 40.24, 13.00, and 5.73 μg/mL, respectively. TDH treatment significantly induced apoptosis and suppressed the viability and migration of human colon cancer LoVo cells. The effect of TDH on induction of apoptosis and inhibition of migration in LoVo cells decreased significantly after activating the PI3K/AKT/mTOR and Wnt/β-catenin signaling pathways with agonists. Additionally, the expression of GOLPH3 protein downregulated significantly in LoVo cells under TDH treatment. Overexpression of the GOLPH3 gene increased the expression of key proteins in PI3K/AKT/mTOR and Wnt/β-catenin signaling pathways and blocked the antitumor activity of TDH. Conclusion. Collectively, the present results indicated that TDH can inhibit the proliferation vitality of colon cancer LoVo cells through downregulating GOLPH3 expression and activity of PI3K/AKT/mTOR and Wnt/β-catenin signaling pathways.

Golgi phosphoprotein 3 induces autophagy and epithelial–mesenchymal transition to promote metastasis in colon cancer

“…Given the role of GOLPH3 in promoting autophagy, we hypothesized that GOLPH3 suppresses Akt, which contradicts the conventional conception that GOLPH3 promotes Akt [ 29 ]. Interestingly, we confirmed our hypothesis and showed that GOLPH3 inhibited Akt phosphorylation at Ser473; this contradicts the majority of previous studies, which reported that GOLPH3 activates the Akt/mTOR signaling to accelerate cancer progression [ 40 , 58 , 59 ]. Previous studies have confirmed that mTOR is a major suppressive regulator of the autophagic process and is regulated by starvation, growth factors, and cellular stresses [ 60 ].…”

“…When Golgi apparatus stress is ameliorated, GOLPH3 expression decreases, and the phosphorylation of the PI3K/Akt/mTOR pathway is promoted [ 62 ]. Previous studies that found GOLPH3 promotes Akt in colon cancer were performed using the cell lines HCT-116 [ 40 ], LoVo [ 58 ], and SW620 [ 63 ]. To the best of our knowledge, no study has constructed and used GOLPH3-overexpressing and -silenced cell lines to investigate the impact of GOLPH3 on Akt in HCT-116 and HT-29 cells.…”

Tenacissoside H Induces Autophagy and Radiosensitivity of Hepatocellular Carcinoma Cells by PI3K/Akt/mTOR Signaling Pathway

“… 13 Moreover, TEH also induced apoptosis and reduced the invasion of colon cancer cells through downregulating GOLPH3 expression and activity of PI3K/AKT/mTOR and Wnt/β-catenin signaling pathways. 29 Our results suggest that TEH suppresses the expression of PI3K/Akt/mTOR signaling pathway and promoted autophagy-related proteins and cell apoptosis. And inhibiting PI3K/Akt/mTOR signaling pathway obviously dampened the anti-tumor role of TEH in HCC cells.…”

IKK/NF‐κB and ROS signal axes are involved in Tenacissoside H mediated inhibitory effects on LPS‐induced inflammatory osteolysis

“…18,19 Tenacissoside H (TDH) is a monomeric extract of Marsdenia tenacissima that has been linked to a few pharmacological benefits, such as anti-inflammatory, antioxidant, and anti-tumour characteristics. [20][21][22] TDH was found to inhibit IL-1β, IL-6 and TNF-α expression and to mediate nitric oxide synthase, as well as inflammation and oxidative stress, thus modifying neurological recovery in mice with cerebral ischemia-reperfusion damage. 20 TDH, on the other hand, has not been studied for osteolytic bone diseases.…”

“…Chinese herbal medicines are clinically natural pharmaceuticals with proven clinical efficacy, and when used properly, they have significantly fewer adverse side effects than western medicine 18,19 . Tenacissoside H (TDH) is a monomeric extract of Marsdenia tenacissima that has been linked to a few pharmacological benefits, such as anti‐inflammatory, antioxidant, and anti‐tumour characteristics 20–22 . TDH was found to inhibit IL‐1β, IL‐6 and TNF‐α expression and to mediate nitric oxide synthase, as well as inflammation and oxidative stress, thus modifying neurological recovery in mice with cerebral ischemia–reperfusion damage 20 .…”