Synergistic Anticancer Activity of Combined Use of Caffeic Acid with Paclitaxel Enhances Apoptosis of Non-Small-Cell Lung Cancer H1299 Cells in Vivo and in Vitro

Abstract

Background/Aims: Caffeic acid (CA) is known to possess multiple biological activities including anti-cancer activities. However, the molecular mechanisms underlying these activities in non-small-cell lung cancer (NSCLC) cells are not fully understood. We attempted to clarify whether CA could enhance paclitaxel (PTX)-induced cytotoxicity in H1299 cells. Methods: First, we tested the cytotoxic effects in both H1299 cells and normal human Bease-2b cells by cell proliferation experiments. Next, we use Annexin V/propidium iodide apoptosis analysis and flow cytometric analysis to investigate apoptosis and cell cycle arrest under the treatments mentioned above. To further pinpoint changes in apoptosis, we tested the caspase-associated apoptotic pathway, which involves the activities of caspase-3 and caspase-9. Moreover, apoptosis-related proteins and MAPK pathway proteins were examined by western blot. An H1299 xenograft nude mice model was used to further evaluate the tumor-suppressing effects of CA and PTX in vivo. Results: Combination treatment with low-dose CA and PTX decreased the proliferation of NSCLC H1299 cells but not normal Beas-2b cells. Flow cytometry showed that H1299 cells were arrested in the sub-G1 phase and apoptosis was significantly increased in H1299 cells after CA treatment. Caspase-3 and caspase-9 activities were both increased after CA treatment. Furthermore, CA increased the PTX-induced activation of Bax, Bid, and downstream cleaved PARP, and phosphorylation of extracellular signal regulated kinase1/2 and c-Jun NH2-terminal protein kinase1/2. An in vivo tumor-suppression assay demonstrated that CA and PTX combined treatment exerted a more effective suppressive effect on tumor growth in H1299 xenografts without causing significant adverse effects. Conclusions: Our results indicated that CA inhibited NSCLC H1299 cell growth by inducing apoptosis and CA and PTX combined produced a synergistic anti-cancer effect in H1299 cells.

Anti-proliferative and apoptosis-inducing effects of matricin on human non-small cell lung cancer H1299 cells via MAPK pathway activation

“…21 Increased expressions of phosphorylated JNK, ERK 1/2, and p-38 proteins with increasing concentration of matricin in the western blot analysis proved that matricin triggered apoptosis in H1299 cells through activation of MAPK pathway. The results of MAPK pathway activation are similarly described in the findings of Min et al 17 Overall, matricin has shown significant anti-proliferative and apoptosis-inducing effects in non-small cell lung cancer H1299 cells. Limitations in this study are the detailed mechanism of apoptosis inducing and the specific small molecule markers responsible for apoptosis were not determined.…”

“…The western blot analysis was performed according to the protocols of Min et al 17 The H1299 cells were pre-treated with matricin (0-80 µM) for 24 h, harvested, and thoroughly rinsed with cold PBS, and centrifuged to obtain cell lysate proteins. Proteins were extracted in radioimmunoprecipitation assay (RIPA) buffer, and immunoblotting was performed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel electrophoresis.…”

Caffeic acid suppresses HT-29 cell death induced by H2O2 via oxidative stress and apoptosis

“…Even though, preceding researches have indicated the effi cacy of CA in inhibiting the viability of cancer cells (Cai et al, 2004, Ishida et al, 2018, Min et al, 2018, this study has pointed out the infl uences of CA in H 2 O 2 -treated HT-29 cells probably to go back to the mismatching results in cancer studies. Davison et al, (2013) and Park (2013) independently reported a controversial information on the infl uence of antioxidant supplement on cancerous cells which was in confl ict with the outdated notion that antioxidants ought to serve as a remedy that would decrease the risk for cancer development by scavenging cancer-leading radicals.…”

Beetroot, A Remarkable Vegetable: Its Nitrate and Phytochemical Contents Can be Adjusted in Novel Formulations to Benefit Health and Support Cardiovascular Disease Therapies

“…Caffeic acid displays antioxidant and anticancer activities [ 123 , 124 , 125 , 126 ], protecting several organs as lungs [ 127 , 128 ], mouth [ 129 , 130 ], liver [ 131 , 132 ], and colon [ 133 , 134 , 135 ]. Caffeic acid can impair macromolecule damage, balancing oxidative stress conditions, and its high antioxidant activity can be ascribed to the hydroxyl groups and the ortho-dihydroxyl group in the caffeic acid molecule [ 136 ].…”