Synergistic antitumor effects via apoptosis of quercetin in combination with cisplatin in the prostate cancer cells / 中华泌尿外科杂志

Objective To study the synergistic antitumor effects of quercetin and cisplatin in human prostate cancer PC3 cells and LNCaP cells.Methods Twelve h after PC3 cells or LNCaP cells were seeded,different dose of quercetin (10 μmol/L,20 μmol/L,40 μmol/L,80 μmol/L,160 μmol/L) or cisplatin (0.01 μmol/L,0.05 μmol/L,0.1 μmol/L,1 μmol/L,10 μmol/L),or quercetin (20 μmol/L) + cisplatin (0.01 μmol/L,0.05 μmol/L,0.1 μmol/L,1 μmol/L,10 μmol/L) were added for48 h and then the antiproliferative effects were detected with MTT assay.After incubated with quercetin (20 μmol/L) or cisplatin (0.05 μ mol/L),or quercetin (20 μ mol/L) + cisplatin (0.05 μmol/L) for48 h,cell cycle distribution and apoptosis of PC3 cells or LNCaP cells were detected by flow cytometer,PI and Annexin V staining.Protein expression was detected by Western blotting.Results After treatment with quercetin or cisplatin alone,the IC50 were (0.99 ± 0.13) μmol/L,(0.75 ± 0.09) μmol/L and (91.60 ± 6.10) μ mol/L,(72.90±4.70) μ mol/L for LNCaP cells or PC3 cells,respectively;The IC50 were (0.11±0.06)μ mol/L,(0.07±0.02) μmol/L for quercetin + cisplatin treatment (Compared with quercetin,P＜0.01 ;Compared with cisplatin,P＜0.05.After treatment with cisplatin or quercetin + cisplatin for 48 h,the S phase percent of LNCaP cells or PC3 cells were (22.4±2.7)％,(31.2±2.4)％ and (20.1±1.6)％,(31.0±2.5)％,respectively,(Compared with control,P＜0.05,however,treatment with quercetin alone has no significant difference (Compared with control,P＞0.05).After treatment with cisplatin or quercetin + cisplatin for 48 h,the apoptotic percent of LNCaP cells or PC3 cells were (14.8 ± 1.9) ％,(39.6 ± 3.1) ％ and (11.5± 1.2) ％,(34.1 ±3.3) ％,respectively,(compared with control,P ＜ 0.05,however,treatment with quercetin alone had no significant difference (compared with control,P＞0.05).After treatment with quercetin alone for 48 h,the activation of caspase-3,caspase-8 and caspase-9 were slightly increased,the expressions of Bax and p21 were up-regulated,the expressions of Bcl-2 and CDK2 were down-regulated.Furthermore,these effect of cisplatin and quercetin + cisplatin were significantly enhanced (compared with quercetin,P＜0.05;compared with quercetin,P＜0.01,respectively.Conclusions The combination modality with quercetin and cisplatin has a better treatment effect in vitro not only in androgen-dependent LNCaP cells but also in androgen-independent PC3 cells.

Molecular mechanism of ophiopogonin B induced cellular autophagy of human cervical cancer HeLa cells / 药学学报

This study is to investigate the antitumor activity of ophiopogonin B (OP-B). MTT assay, flow cytometric analysis, acridine orange staining, Lyso-Tracker Red staining and HeLa-GFP-LC3 transfect cells assay were used to detect the proliferation activity, apoptosis and autophagy of HeLa cells. The results showed that OP-B exerted potent antiproliferative activity on HeLa cells, the cell growth inhibition effect of OP-B was not due to apoptosis and OP-B could induce autophagy of HeLa cells. OP-B also induced the protein expression up-regulation of Beclin-1 and promoted LC3 I transformation LC3 II, which were representative proteins of autophagy. Furthermore, 3-MA, an inhibitor of autophagy, not only inhibited OP-B-mediated autophagy but also almost completely reversed the antiproliferative effect of OP-B, suggesting that the growth inhibition effect of OP-B was autophagy dependent. Western blotting demonstrated that OP-B inhibited the phosphorylation of Akt and its' downstream vital protein, such as mTOR and p70S6K. In addition, OP-B also induced the protein expression up-regulation of PTEN, which is a negative regulation protein for Akt/mTOR signaling pathway. However, OP-B did not affect the protein expression of total Akt. Collectively, the antitumor effects of OP-B were autophagy-dependent via repression Akt/mTOR signaling pathway. Therefore, OP-B is a prospective inhibitor of Akt/mTOR and may be used as an alternative compound to treat cervical carcinoma.

Synergistic antitumor effects of tanshinone II A in combination with cisplatin via apoptosis in the prostate cancer cells / 药学学报

Treatment with the combination of Chinese herbs and cytotoxic chemotherapies showed a higher survival rate in clinical trials. In this report, the results demonstrated that the tanshinone II A, a key component of Salvia miltiorrhiza bunge, when it is combined with the cytotoxic drug cisplatin showed synergistic antitumor effects on human prostate cancer PC3 cells and LNCaP cells in vitro. Antiproliferative effects were detected with MTT assay. Cell cycle distribution and apoptosis were detected by flow cytometer. Protein expression was detected by Western blotting. The intracellular concentration of cisplatin was detected by high performance liquid chromatography. The results demonstrated that tanshinone II A significantly enhanced the antiproliferative effects of cisplatin on human prostate cancer PC3 cells and LNCaP cells with the increase of the intracellular concentration of cisplatin. These effects were correlated with cell cycle arrested at S phase and cell apoptosis. The apoptosis might be achieved through death receptor pathway and mitochondrial pathway. Furthermore, the Bcl-2 family members were also involved in this apoptotic process. Collectively, these results indicated that the combination of tanshinone II A and cisplatin had a better treatment effect in vitro not only on androgen-dependent LNCaP cells but also on androgen-independent PC3 cells.

NNIspm, a polyamine derivative, induces cellular senescence of human hepatoma HepG2 cells and its molecular mechanism / 药学学报

This study is to examine the effects of NNIspm-mediated cellular senescence of HepG2 cells and elucidate its potential molecular mechanism. Cellular senescence was detected with senescence-associated beta-galactosidase staining. Cell cycle distribution, intracellular fluorescence intensity and accumulation of intracellular reactive oxygen species (ROS) were detected by high content screening (HCS). Protein expression was detected by Western blotting. Polyamines content was analyzed by high performance liquid chromatography (HPLC). The results demonstrated that NNIspm significantly induced HepG2 cells senescence. This effect was due to the decrease of intracellular polyamines, the arrest at G0/G1 phase and an increase of ROS level. The molecular senescence marker p21 increased significantly after NNIspm treatment. In contrast, the protein expressions of Cyclin E and CDK2 were obvious down-regulation. The results indicated that cellular senescence induced by NNIspm was one of its antitumor mechanisms.

Part IV. Synthesis and antitumor evaluation of s-triazolothiadiazines and pyrazolo s-triazoles derived from ciproxacin / 药学学报

An efficient modified route based on the targeting mechanism of antibacterial fluoroquinolones for the shift from the antibacterial activity to the antitumor one was further developed. Using a fused heterocyclic ring, s-triazolothiadiazine as a carboxyl bioisostere of ciprofloxacin, the title compounds, 1-cyclopropyl-6-fluoro-7-piperazin-1-yl-3-(6-substituted-phenyl-7H-[1, 2, 4]triazolo[3, 4-b][1, 3, 4]thiadiazin-3-yl)-quinolin-4(1H)-ones (5a-5e) and their corresponding N-acetyl products (6a-6e), were designed and synthesized, separately. Meaningfully, a ring-contraction of fused six-membered thiadiazine occurred by a sulfur extrusion reaction gave new tri-acetylated fused heterocycles related to pyrazolo[5, 1-c][1, 2, 4] triazoles (7a-7e). The in vitro antitumor activity against L1210, CHO and HL60 cell lines was also evaluated for the synthesized fifteen heterocycles compared to parent ciprofloxacin by methylthiazole trazolium (MTT) assay. Interestingly, the results displayed that fifteen fused heterocyclic compounds showed more significant growth inhibitory activity (IC50 150.0 micromol x L(-1)), and the active order decreased from 7a-7e to 5a-5e to 6a-6e, respective.

Antiproliferative effects mechanism of beta-sitosterul in hepatoma HepG2 cells / 中国中药杂志

<p><b>OBJECTIVE</b>To study the antiproliferative effects of beta-sitosterul and its mechanism in hepatoma HepG2 cells.</p><p><b>METHOD</b>Cell proliferation was assessed by MTT assay. Cell cycle distribution, apoptosis and mitochondrial membrane potential were measured by high content screening (HCS). The protein expression of caspase-3, caspase-8, caspase-9, Bcl-2, Bax, tBid and cytochrome c in the HepG2 cells were evaluated by Western Blots.</p><p><b>RESULT</b>beta-Sitosterul exerted significant antiproliferative effects in HepG2 cells. Furthermore, beta-sitosterul also induced HepG2 cells apoptosis, lost mitochondrial membrane potential, activated caspase-3, caspase-8 and caspase-9, up-regulate Bax, tBid protein, down-regulation Bcl-2 protein. However, beta-sitosterul had hardly any effects on QSG7701 cells.</p><p><b>CONCLUSION</b>beta-Sitosterul exerted antiproliferative effects and induced HepG2 cells apoptosis via mitochondrial pathway and membrane death receptor pathway.</p>

Involvement of p38-p53 signal pathway in resveratrol-induced apoptosis in MCF-7 cells / 药学学报

This paper is to report the study of resveratrol-induced apoptosis and its mechanisms in MCF-7 cells. MTT assay was performed to assess the cytotoxicity of resveratrol on MCF-7 cells. Hoechst 33258 staining was used to observe cellular morphologic changes in apoptosis. Apoptosis was measured by flow cytometric analysis and the protein expression was examined by Western blotting analysis. The results indicated that resveratrol could inhibit MCF-7 cell growth in a time- and concentration-dependent manner. Remarkable morphologic changes in the cells after 60 micromol L(-1) resveratrol treatment, including cell nuclear shrinkage, DNA condensation and apoptotic bodies, were observed by Hoechst 33258 staining. Resveratrol could induce apoptosis and activate p38 and p53 in a time dependent manner in MCF-7 cells. In addition, the cell growth inhibitory ratio and the apoptotic ratio of resveratrol-treated group decreased markedly by the p38 MAPK inhibitor SB203580 or p53 inhibitor pifithrin-alpha. Further experiments confirmed that resveratrol-induced p53 activation was reduced by SB203580 whereas the activation of p38 was not affected by pifithrin-alpha. In conclusion, resveratrol induced apoptosis in MCF-7 cells could be through activating p38-p53 signal pathway.

Acetylsalicylic acid strengthens the effects of ANISpm against hepatocellular carcinoma and its molecular mechanism / 药学学报

The objective of this study is to examine the effects of ANISpm, a novel polyamine naphthalimide conjugate, with acetylsalicylic acid against hepatocellular carcinoma in vivo and in vitro and elucidate its potential molecular mechanism. The proliferation inhibition was detected by MTT assay. Cell apoptosis, intracellular fluorescence intensity and mitochondrial membrane potential (MMP) were detected by high content screening (HCS) analysis. Polyamines content was analyzed by reverse-phase high performance liquid chromatography Protein expression levels were quantified by Western blotting assay. The combination treatment strongly inhibited cell proliferation, induced cell apoptosis in HepG2 cells and H22 hepatoma cells, which was mediated by enhanced ANISpm uptake via up-regulation of spermidine/spermine N1-acetyltransferase (SSAT) and depression of intracellular polyamine. Furthermore, this synergistic apoptosis was involved in mitochondria and death-receptor signal pathway. All these findings demonstrated that the combination treatment with acetylsalicylic acid and ANISpm resulted in synergistic antitumor effects on hepatoma cells. Thus, combination therapy with these agents may be useful as a potential template for the development of better chemotherapeutic strategy against hepatoma.

NNINspm,naphthalimide-polyamine conjugate,induces hepatoma HepG2 cell apoptosis via PI3K/Akt pathways / 中国药理学通报

Aim To investigate the apoptotic mechanism and polyamine transporter recognition of 3-nitro-naphthalimide norspermine conjugate (NNINspm),a novel naphthalimide-polyamine conjugate, in HepG2 cells.Methods The cytotoxicity of NNINspm was assessed by MTT assay.Cell cycle distribution and apoptosis were measured by flow cytometry.The protein expression of cytochrome C,14-3-3,Bad,Bcl-xL,mTOR,p70S6K,Cdk4,p27~(kip1),Akt,Caspase-3,Caspase-9 was evaluated by Western blot.The translocation of Akt was detected by high content screening (HCS) analysis.Results NNINspm induced HepG2 cells apoptosis via Akt dephosphorylation and then triggered a series of signal events, such as Bad dephosphorylation, dissociation of 14-3-3 and Bad, and then binding to Bcl-xL,which finally resulted in mitochondrial disruption,cytochrome c release and caspase cascade activation.Furthermore,the NNINspm-mediated cell cycle arrest was due to mTOR and p70S6K dephosphorylation,Cdk4 down-regulation and p27~(kip1) up-regulation.Conclusion NNINspm induces HepG2 cell apoptosis via PI_3K/Akt signal pathway.

Study on the action mechanism for δ-tocotrienol-induced apoptosis of human hepatoma HepG2 cells / 肿瘤

Objective:To elucidate action mechanism of δ-tocotrienol in inducing apoptosis of human hepatoma HepG2 cells. Methods:Cell proliferation and viability were assessed by MTT assay; cell cycle distribution, apoptotic rate and mitochondrial membrane potential were measured by using high content screening system; the expression of apoptosis-related protein such as caspase-3, caspase-8, caspase-9, Bcl-2, Bax, tBid and cytochrome C in the HepG2 cells were evaluated by Western blotting. Results:δ-Toco-trienol inhibited HepG2 cell proliferation and induced apoptosis in a dose-dependent manner. This growth-inhibiting effect of δ-toco-trienol correlated with loss of mitochondrial membrane potential and release of cytochrome C from mitochondria to cytoplasm, and regulation of the protein expression of Bcl-2 family members, such as up-regulation of Bax and tBid and down-regulation of Bcl-2. Subsequently tocotrienol induced the activation of caspase-3, caspase-8, and caspase-9 which finally induced apoptosis of hepatoma HepG2 cells. Conclusion:δ-Tocotrienol induced apoptosis of human hepatoma HepG2 cells via mitochondrial pathway and membrane death receptor pathway.

Part II: Design, synthesis and antitumor action of C3/C3 bisfluoroquinolones linked-cross 2, 5-1, 3, 4oxadiazole / 药学学报

To develop a new small molecular probe for discovering an antitumor lead compound from the replacement of carboxylic group of two molecular antibacterial fluoroquinolones with a heterocyclic ring, a series of the C3/C3 bis-fluoroquinolones tethered with an 1, 3, 4-oxadiazole ring were synthesized as their respective HCl salts, and their structures were characterized by elemental analysis and spectral data. The in vitro antitumor activity against L1210, CHO and HL60 cell lines was also evaluated via the respective IC50 values by methylthiazole trazolium (MTT) assay.

Apoptotic mechanism of WJH-6, a novel polyamine conjugate, on K562 and HL-60 cells / 药学学报

In the present study, the apoptotic mechanism and polyamine transporter recognition of WJH-6, a novel polyamine conjugate, were investigated in K562 and HL-60 cells. The cytotoxicity of WJH-6 was assessed by MTT assay; cell cycle distribution and apoptosis were measured by flow cytometry; the protein expression of Caspase-3, Caspase-8, Caspase-9, Bid and mitochondrial membrane potential (MMP) were evaluated by high content screening (HCS) analysis; the protein expression of cytochrome c was measured by Western blotting. The results showed that WJH-6 could be recognized and transported by polyamine transporter (PAT). Furthermore, WJH-6 was able to inhibit K562 and HL-60 cells proliferation and induce apoptosis. This apoptotic effect was relative to MMP loss, cytochrome c release from mitochondria to cytoplasm and the activation of Caspase-8, Caspase-9, Caspase-3 and Bid. These results suggested that WJH-6-induced K562 and HL-60 cells apoptosis was related with mitochondrial damage.

Antitumor mechanism of HG251,a novel anthracene derivative,in K562/DOX leukemia cells / 中国药理学通报

Aim To evaluate the mechanism of HG251-induced apoptosis in K562/DOX cells.Methods Cell viability was assessed by MTT assay;cell cycle distribution,apoptosis and mitochondrial membrane potential were measured by flow cytometry;the protein expressions of P-gp,caspase-3,caspase-8,caspase-9,p53,Bcl-xL and cytochrome c in the K562/DOX cells were evaluated by Western blot.Results HG251 was able to inhibit cells proliferation,induce apoptosis,lose mitochondrial membrane potential,activate caspase-3,caspase-8,caspase-9,up-regulate p53 protein and down-regulate Bcl-xL protein in a dose-dependent manner but it had no effect on P-gp protein expression.Conclusion HG251 could overcome apoptotic resistance via up-regulating p53 protein and down-regulating Bcl-xL protein.In addition,HG251 also induced K562/DOX cells apoptosis via mitochondrial pathway and membrane death receptor pathway.

NNAMB,a novel homospermidine conjugate,induces apoptosis and differentiation in B16 Melanoma cells / 中国药理学通报

0.1 ?mol?L-1), inducing differentiation through enhancement of melanogenesis and increase of the activity of tyrosinase at lower doses(