Hederagenin from the leaves of ivy (Hedera helix L.) induces apoptosis in human LoVo colon cells through the mitochondrial pathway

Abstract

BackgroundColorectal cancer has become one of the leading cause of cancer morbidity and mortality throughout world. Hederagenin, a derivative of oleanolic acid isolated from the leaves of ivy (Hedera helix L.), has been shown to have potential anti-tumor activity. The study was conducted to evaluate whether hederagenin could induce apoptosis of human colon cancer LoVo cells and explore the possible mechanism.MethodsMTT assay was used for evaluating cell viability while Annexin V-FITC/PI assay and Hoechst 33342 nuclear stainining were used for the determination of apoptosis and mitochondrial membrane potential. DCFH-DA fluorescence staining and flow cytometry were used to measure ROS generation. Real-time PCR and western blot analysis were performed for apoptosis-related protein expressions.ResultsMTT assay showed that hederagenin could significantly inhibit the viability of LoVo cells in a concentration-dependent and time-dependent manner by IC50 of 1.39 μM at 24 h and 1.17 μM at 48 h. The apoptosis ratio was significantly increased to 32.46% and 81.78% by the induction of hederagenin (1 and 2 μM) in Annexin V-FITC/PI assay. Hederagenin could also induce the nuclear changes characteristic of apoptosis by Hoechst 33342 nuclear stainining under fluorescence microscopy. DCFH-DA fluorescence staining and flow cytometry showed that hederagenin could increase significantly ROS generation in LoVo cells. Real-time PCR showed that hederagenin induced the up-regulation of Bax and down-regulation of Bcl-2, Bcl-xL and Survivin. Western blotting analysis showed that hederagenin decreased the expressions of apoptosis-associated proteins Bcl-2, procaspase-9, procaspase-3, and polyADP- ribosepolymerase (PARP) were increased, while the expressions of Bax, caspase-3, caspase-9 were increased. However, there was no significant change on caspase-8.ConclusionsThese results indicated that the disruption of mitochondrial membrane potential might contribute to the apoptosis of hederagenin in LoVo cells. Our findings suggested that hederagenin might be a promising therapeutic candidate for human colon cancer.

125 I seeds irradiation inhibits tumor growth and induces apoptosis by Ki-67, P21, Survivin，Livin and Caspase-9 expression in lung carcinoma xenografts

“…The central event in apoptosis is the proteolytic activation of a class of cysteine aspartyl-specific proteases(Caspase) family (38,45,(50)(51)(52)(53)(54)(55). Caspases are known to act as important mediators of apoptosis and contribute to the overall apoptotic morphology by the cleavage of various cellular substrates (44,49,54).…”

“…This gene encodes a protein that belongs to a highly conserved family of cysteinyl aspartate-specific proteases that function as essential regulators of programmed cell death through apoptosis. The increase of caspase-3 expression may induce the mitochondria-dependent apoptotic pathway with the possible mitochondria-independent pathway for the caspase-8 activation for that caspase-8-mediated apoptosis induced by oxidative stress is independent of the intrinsic pathway and dependent on cathepsins (38,45,(50)(51)(52)(53)(54)(55). The intrinsic pathway of apoptosis is associated with the activation of caspase-9, which cleaves and activates caspase-3 (49)(50)(51)(52)(53)(54).…”

Current knowledge and development of hederagenin as a promising medicinal agent: a comprehensive review

“…4 In recent years, HG has been observed to be biologically active, as well as many other saponins with similar structures, such as ursolic acid and corosolic acid. HG exhibits multiple pharmaceutical and biological activities, including antitumor, 2,[5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23] anti-inammatory, [24][25][26][27][28][29][30] anti-depressant, [31][32][33][34][35] antineurodegenerative, [36][37][38] anti-hyperlipidemia, 4,39,40 anti-diabetic, [41][42][43] anti-leishmanial, 19,44,45 and anti-viral (against hepatitis B) activity. 46 Recently, a number of studies have investigated the use of HG as a potential anti-tumor agent.…”

“…46 Recently, a number of studies have investigated the use of HG as a potential anti-tumor agent. 5,6,[10][11][12][14][15][16] The pharmacokinetics, [47][48][49][50][51] structural modication, [17][18][19][20][21]35,38,45,46 and mechanisms of action [4][5][6][10][11][12][13][14][15][16]19,25,26,[31][32][33][34]36,37,39,40,43,45 of HG have also been studied. However, possibly owing to limited in vivo data, HG is still in the preclinical development stage.…”

Luteolin induces myelodysplastic syndrome‑derived cell apoptosis via the p53‑dependent mitochondrial signaling pathway mediated by reactive oxygen species

“…Kittiratphatthana et al (17) reported that luteolin exerts its pro-apoptotic effect partly through generating intracellular ROS, which then contributes to the induction of mitochondria-mediated cell apoptosis. Others have indicated that the overexpression of ROS may contribute to the apoptosis through regulating B-cell lymphoma 2 (Bcl-2) family proteins, releasing cytochrome c from the mitochondria, and activating effector caspases, including caspase-3 and caspase-9 (21). Therefore, a promising strategy for the prevention and treatment of MdS has emerged via inducing apoptosis by mitochondria-mediated ROS generation.…”