α­Solanine inhibits growth and metastatic potential of human colorectal cancer cells.

Solanum nigrum L. (Longkui) is one the most widely used anticancer herbs in traditional Chinese medicine. α­Solanine is an important ingredient of S. nigrum L. and has demonstrated anticancer properties in various types of cancer. However, the effects of α­solanine on colorectal cancer remain elusive. The aim of the present study was to assess the effects of α­solanine on human colorectal cancer cells. The results demonstrated that α­solanine inhibited the proliferation of RKO cells in a dose­ and time­dependent manner. In addition, α­solanine arrested the cell cycle at the G0/G1 phase and suppressed the expression levels of cyclin D1 and cyclin­dependent kinase 2 in RKO cells. α­Solanine induced apoptosis of RKO cells, as indicated by morphological changes and positive Annexin­FITC/propidium iodide staining. Additionally, α­solanine activated caspase­3, ­8 and ­9 in RKO cells, which contributed to α­solanine­induced apoptosis. α­Solanine also increased the generation of reactive oxygen species, which contributed to caspase activation and induction of apoptosis. α­Solanine inhibited the migration, invasion and adhesion of RKO cells, as well as the expression levels and activity of matrix metalloproteinase (MMP)­2 and MMP­9. In addition, α­solanine inhibited cell proliferation, activated caspase­3, ­8 and ­9, induced apoptosis, and inhibited the migration and invasion of HCT­116 cells. Furthermore, α­solanine inhibited tumor growth and induced apoptosis in vivo. These findings demonstrated that α­solanine effectively suppressed the growth and metastatic potential of human colorectal cancer.

Weichang'an Formula Inhibits Tumor Growth in Combination with Bevacizumab in a Murine Model of Colon Cancer-Making up for the Deficiency of Bevacizumab by inhibiting VEGFR-1.

Aim: Angiogenesis plays an important role in the initiation, development, and metastasis of malignant tumors. Antiangiogenic drugs combined with immune therapy are considered to have a synergistic effect on anti-tumor strategy. Weichang'an formula (WCAF) is a prescription of traditional Chinese medicine (TCM) based on pharmaceutical screening and clinical experience. The aim of this study is to examine the effect of WCAF and its combined action with Bevacizumab (BEV) in colorectal cancer, and to identify the possible mechanism of action. Methods: A human colon cancer cell (HCT 116) subcutaneous xenograft model was established in BALB/c-nu/nu mice. Tumor-bearing mice were randomized into each of four groups: control, WCAF treated, BEV treated, and WCAF plus BEV treated. Apoptosis was detected by TUNEL assay. Western blot was used to assess the protein levels of Leptin-R, STAT3, p-STAT3, BCL-2, and VEGFR-1. Immunohistochemistry was used to detect the micro-vessel density (MVD) and AKT1. Leptin and Vascular endothelial growth factor A (VEGF-A) mRNA expression were detected by Real-time PCR (RT-PCR). A network pharmacology study and validation assay were carried out to find the underlying molecular targets of WCAF related to immune regulation. Results: Compared with the control group, WCAF reduced tumor weight and volume, as well as promoted tumor cell apoptosis. WCAF treatment decreased the mRNA expression of Leptin and VEGF-A, while the protein levels of CD31, LEP-R, VEGFR-1, STAT3, and p-STAT3 were decreased in tumor tissues. In addition, VEGFR-1 protein expression was decreased in the WCAF group and the WCAF plus BEV group but not in the BEV group. The combination of WCAF and BEV demonstrated a partial additive anti-tumor effect in vivo. The pharmacological network also found there are 26 WCAF target proteins related to cancer immune and 12 cancer immune related pathways. The AKT1 protein expression in the WCAF and WCAF + BEV groups were significantly lower than the that in the control group (p < 0.01). Conclusion: WCAF can inhibit tumor growth and promote apoptosis and inhibit tumor angiogenesis in subcutaneous xenografts of human colon cancer HCT-116 in nude mice. WCAF also makes up for the deficiency of BEV by inhibiting VEGFR-1. The VEGFR-1 expression between the combination group and BEV alone achieved statistically significant difference (p < 0.01). Combined with BEV, WCAF showed a partial additive anti-tumor effect. The mechanism may be related to Leptin/STAT3 signal transduction, VEGF-A, VEGFR-1 and WCAF target proteins related to cancer immune such as leptin and AKT1.

Eucommia ulmoides Oliv. Leaf Extract Improves Erectile Dysfunction in Streptozotocin-Induced Diabetic Rats by Protecting Endothelial Function and Ameliorating Hypothalamic-Pituitary-Gonadal Axis Function.

Erectile dysfunction (ED) is a major complication of diabetes mellitus. Eucommia ulmoides Oliv. is used as a traditional medicine for male impotence, but no systematic study has examined its effect on diabetes-associated ED. In this study, we investigated the effects of Eucommia ulmoides Oliv. leaf extract (EULE) on restoring erectile function in streptozotocin (STZ)-induced diabetic rats model. After 16 weeks of treatment, EULE administration had significantly increased intracavernosal pressure, nitric oxide (NO) levels, and cyclic guanosine monophosphate (cGMP) concentrations. Serum superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels were markedly higher and serum malondialdehyde (MDA) levels were lower in the EULE-treated groups than in the diabetic model group. EULE restored NO biosynthesis by significantly increasing protein kinase B (Akt) and endothelial NO synthase (eNOS) activation. Furthermore, EULE is likely to benefit the hypothalamic-pituitary-gonadal (HPG) axis, as it increased gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T) concentrations as well as hormone receptors Gnrhr, Fshr, and Lhr expression levels. Hence, EULE attenuates oxidative stress, increases NO production, and activates the Akt-eNOS pathway to restore endothelial function; moreover, EULE enhances the HPG axis to improve erectile function. These results suggest that EULE may represent a new therapeutic avenue for diabetes-associated ED.

Combined administration of anisodamine and neostigmine rescued acute lethal crush syndrome through α7nAChR-dependent JAK2-STAT3 signaling.

Previously we showed that Ani (anisodamine)/Neo (neostigmine) combination produced anti-shock effect via activating α7 nicotinic acetylcholine receptor (α7nAChR). In this study, we aim to investigate the therapeutic effect and underlying mechanisms of Ani/Neo combination in acute lethal crush syndrome (CS). In rat and rabbit CS models, Ani/Neo combination increased the 24 h survival rates, improved hemodynamics and decreased the levels of creatine kinase, MB isoenzyme of creatine kinase, blood urea nitrogen, creatinine, K+ in serum. It also decreased the levels of H2O2, myeloperoxidase (MPO) and nitric oxide (NO) in serum and compressed muscle in rat CS model. In wild-type (WT) mice with CS, Ani/Neo combination increased 24 h survival rate and decreased the levels of H2O2, MPO, NO, TNFα, IL-6 and IL-10 in compressed muscle. These effects were attenuated by α7nAChR knockout (KO). Moreover, Ani/Neo combination prevented the decrease of phosphorylation of Janus kinase 2 (JAK2) and phosphorylation of signal transducer and activator of transcription 3 (STAT3) induced by CS. These effects of Ani/Neo in CS mice were cancelled by methyllycaconitine (α7nAChR antagonist) and α7nAChR KO. Collectively, our results demonstrate that Ani/Neo combination could produce therapeutic effects in CS. The underlying mechanism involves the activation of α7nAChR-dependent JAK2-STAT3 signaling pathway.

Activation of phenylpropanoid pathway and PR of potato tuber against Fusarium sulphureum by fungal elicitor from Trichothecium roseum.

The induced resistance of potato tuber (Solanum tuberosum cv. Xindaping) tissue against Fusarium sulphureum by a fungal elicitor from the incompatible pathogen Trichothecium roseum and its possible mechanism were studied. The results showed that the lesion development of the wound-inoculated potato tuber was significantly reduced by treatment with the fungal elicitor from T. roseum (P < 0.05). Inoculation with F. sulphureum on the 16th day after treatment with the fungal elicitor80 at 15.0 µg/ml had the best resistant effect in the potato tuber, with the diameter being only reduced by 47 % that of the control. In addition, the results also showed that the potato tuber treated with the fungal elicitor80 could systemically induce lignin deposition, total phenolic content, flavonoid content and defense enzymes, including three keys phenylpropanoid pathway (PAL, 4CL and C4H) and pathogenesis-related (GLU and CHT) enzymes. The fungal elicitor80 also enhanced the up-regulation of the transcription and expression of PAL, C4H, 4CL, GLU and CHT genes. The treatment with the fungal elicitor80 + F. sulphureum caused the marked and/or prompt enhancement of all indexes when compared to treatment with the fungal elicitor80 or inoculation with the pathogen alone. The results suggested that the fungal elicitor of T. roseum could significantly enhance defense responses in potato tuber against dry rot mainly due to the up-regulation of the transcription and expression of resistance-related genes as well as increasing the activity of resistance-related enzymes and antifungal compounds.

[Research strategy of traditional Chinese medicine proteomics].

The application of proteomics in the research of traditional Chinese medicine (TCM) is very extensive, and there have been many successful cases. In this paper, the previous studies on the complex system of TCM by using proteomics technology were reviewed, and the authors proposed to set up a special subject on proteomics in TCM, which is called TCM proteomics. In this paper, the research strategies and the future research directions of TCM proteomics were reviewed and discussed, which may provide some ideas for the researchers of TCM proteomics.

Research strategy of traditional Chinese medicine proteomics / 中国中药杂志

The application of proteomics in the research of traditional Chinese medicine (TCM) is very extensive, and there have been many successful cases. In this paper, the previous studies on the complex system of TCM by using proteomics technology were reviewed, and the authors proposed to set up a special subject on proteomics in TCM, which is called TCM proteomics. In this paper, the research strategies and the future research directions of TCM proteomics were reviewed and discussed, which may provide some ideas for the researchers of TCM proteomics.

Ligustrum lucidum Ait. fruit extract induces apoptosis and cell senescence in human hepatocellular carcinoma cells through upregulation of p21.

Nü-zhen-zi, the fruit of Ligustrum lucidum Ait., is one of the most frequently used liver Yin tonifying Chinese herbs for the treatment of liver cancer. However, the effect of Ligustrum lucidum fruit on hepatocarcinoma cells remains unknown. In the present study, we evaluated the effects of a Ligustrum lucidum fruit extract (LLFE) on human hepatocellular carcinoma Bel-7402 cells. The results showed that LLFE inhibited the proliferation of the Bel-7402 cells in a dose- and time-dependent manner. LLFE induced apoptosis in Bel-7402 cells accompanied by activation of caspase-3, -8 and -9. LLFE-induced apoptosis was completely abrogated by a pan caspase inhibitor, Z-VAD-FMK. LLFE treatment also caused a large and flat morphologic cellular change, positive SA-ß-gal staining, and G0/G1 phase cell cycle arrest in the Bel-7402 cells, accompanied by upregulation of p21 and downregulation of RB phosphorylation. Specific knockdown of p21 expression by RNA interference partially abrogated LLFE-induced apoptosis, and significantly abrogated LLFE-induced cell senescence. These observations suggest that Nü-zhen-zi is a potential anticancer herb and support the traditional use of Nü-zhen-zi for hepatocarcinoma treatment.

[Controlled observation of the efficacy between floating acupuncture at Tianying point and warm-needling therapy for supraspinous ligament injury].

OBJECTIVE: To compare the efficacy difference in the treatment of supraspinous ligament injury between floating acupuncture at Tianying point and the conventional warm needling therapy. METHODS: Ninety patients were randomized into a floating acupuncture group and a warm needling group, 45 cases in each one. In the floating acupuncture group, the floating needling technique was adopted at Tianying point. In the warm needling group, the conventional warm needling therapy was applied at Tianying point as the chief point in the prescription. The treatment was given 3 times a week and 6 treatments made one session. The visual analogue scale (VAS) was adopted for pain comparison before and after treatment of the patients in two groups and the efficacy in two groups were assessed. RESULTS: The curative and remarkably effective rate was 81.8% (36/44) in the floating acupuncture group and the total effective rate was 95.5% (42/44), which were superior to 44.2% (19/43) and 79.1% (34/43) in the warm needling group separately (P < 0.01, P < 0.05). VAS score was lower as compared with that before treatment of the patients in two groups (both P < 0.01) and the score in the floating acupuncture group was lower than that in the warm needling group after treatment (P < 0.01). Thirty-six cases were cured and remarkably effective in the floating acupuncture group after treatment, in which 28 cases were cured and remarkably effective in 3 treatments, accounting for 77.8 (28/36), which was apparently higher than 26.3 (5/19) in the warm-needling group (P < 0.01). CONCLUSION: The floating acupuncture at Tianying point achieves the quick and definite efficacy on supraspinous ligament injury and presents the apparent analgesic effect. The efficacy is superior to the conventional warm-needling therapy.

Teng-Long-Bu-Zhong-Tang, a Chinese herbal formula, enhances anticancer effects of 5--Fluorouracil in CT26 colon carcinoma.

BACKGROUND: Colorectal cancer remains one of the leading causes of cancer death worldwide. Traditional Chinese Medicine (TCM) has played a positive role in colorectal cancer treatment. There is a great need to establish effective herbal formula for colorectal cancer treatment. Based on TCM principles and clinical practices, we have established an eight herbs composed formula for colorectal cancer treatment, which is Teng-Long-Bu-Zhong-Tang (TLBZT). We have demonstrated the anticancer effects of TLBZT against colorectal carcinoma in vitro. In present study, we evaluated the anticancer potential of TLBZT, used alone or in combination with low dose of 5-Fluorouracil (5-Fu), in CT26 colon carcinoma in vivo. METHODS: CT26 colon carcinoma was established in BALB/c mice and treated with TLBZT, 5-Fu, or TLBZT plus 5-Fu. The tumor volumes were observed. Apoptosis was detected by TUNEL assay. Caspases activities were detected by colorimetric assay. Cell senescence was indentified by senescence ß-galactosidase staining. Gene expression and angiogenesis was observed by immunohistochemistry or western blot. RESULTS: TLBZT significantly inhibited CT26 colon carcinoma growth. TLBZT elicited apoptosis in CT26 colon carcinoma, accompanied by Caspase-3, 8, and 9 activation and PARP cleavage, and downregulation of XIAP and Survivin. TLBZT also induced cell senescence in CT26 colon carcinoma, with concomitant upregulation of p16 and p21 and downregulation of RB phosphorylation. In addition, angiogenesis and VEGF expression in CT26 colon carcinoma was significantly inhibited by TLBZT treatment. Furthermore, TLBZT significantly enhanced anticancer effects of 5-Fu in CT26 colon carcinoma. CONCLUSIONS: TLBZT exhibited significantly anticancer effect, and enhanced the effects of 5-Fu in CT26 colon carcinoma, which may correlate with induction of apoptosis and cell senescence, and angiogenesis inhibition. The present study provides new insight into TCM approaches for colon cancer treatment that are worth of further study.

Synergistic anticancer effects of curcumin and resveratrol in Hepa1-6 hepatocellular carcinoma cells.

Hepatocellular carcinoma remains one of the most prevalent malignancies worldwide. Curcuma aromatica and Polygonum cuspidatum are one of the commonly used paired-herbs for liver cancer treatment. Curcumin and resveratrol are the major anticancer constituents of Curcuma aromatica and Polygonum cuspidatum, respectively. Curcumin and resveratrol have been found to exhibit a synergistic anticancer effect in colon cancer. However, the combined effect of curcumin and resveratrol against hepatocellular carcinoma remains unknown. In the present study, we evaluated the combined effects of curcumin and resveratrol in hepatocellular carcinoma Hepa1-6 cells. The results showed that curcumin and resveratrol significantly inhibited the proliferation of Hepa1-6 cells in a dose- and time-dependent manner. The combination treatment of curcumin and resveratrol elicited a synergistic antiproliferative effect in Hepa1-6 cells. The apoptosis of Hepa1-6 cells induced by the combination treatment with curcumin and resveratrol was accompanied by caspase-3, -8 and -9 activation, which was completely abrogated by a pan caspase inhibitor, Z-VAD-FMK. Combination of curcumin and resveratrol upregulated intracellular reactive oxygen species (ROS) levels in Hepa1-6 cells. The ROS scavenger, NAC, partially attenuated the apoptosis and caspase activation induced by the combination treatment of curcumin and resveratrol. In addition, the combination of curcumin and resveratrol downregulated XIAP and survivin expression. These data suggest that the combination treatment of curcumin and resveratrol is a promising novel anticancer strategy for liver cancer. The present study also provides new insights into the effective mechanism of paired-herbs in traditional Chinese medicine.

[Effect of Solanum nigrum on human colon carcinoma RKO cells ].

OBJECTIVE: To evaluate the effect of Solanum nigrum on adhesion, migration and invasion in human colon carcinoma RKO cells. METHODS: RKO cells were treated with different dose of Solanum nigrum. Cell proliferation was detected by CCK-8 assay. Cell adhesion was observed with CytoSelect 48-Well Cell Adhesion Assay. Cell migration was detected with scratch assay. Cell invasion was analyzed by CytoSelect 24-Well Cell Invasion Assay. RESULTS: At final concentration of 400-1600 microg/mL, Solanum nigrum significantly inhibited proliferation of RKO cells in a dose-dependent manner. At final concentration of 100-400 microg/mL, Solanum nigrum significantly inhibited adhesion,migration and invasion in RKO cells. CONCLUSION: Solanum nigrum may inhibit the proliferation, adhesion, migration and invasive abilities in RKO cells. The present study provides new insight into the application of Solanum nigrum for colon carcinoma treatment that are worthy of further study.

Inhibitory effects of Qushuanling Capsule () on thrombus formation and platelet aggregation in rats.

OBJECTIVE: To investigate the effects of Qushuanling Capsule ( QSLC) on thrombus formation and platelet aggregation in rats. METHODS: Arteriovenous bypass, venous thrombosis, and middle cerebral artery thrombosis models were used in rats to investigate the anti-thrombotic effects of QSLC, a compound of nine Chinese herbs. The platelet aggregation induced by adenosine diphosphate (ADP), thrombin or arachidonic acid (AA), as well as the contents of thromboxane B(2) (TXB(2)) and 6-keto-prostaglandin F1α (6-keto-PGF1α) in rat plasma and aortic walls, were determined to investigate the possible mechanisms of the anti-thrombotic effects of QSLC. RESULTS: After oral administration with QSLC for 7 days, arteriovenous bypass thrombosis was obviously suppressed compared with the model group, venous thrombosis was also obviously suppressed, rat behaviors were obviously improved, and brain infarct size as well as water content were also reduced. The platelet aggregation induced by ADP or thrombin was inhibited by QSLC, but the drug had no effect on AA-induced platelet aggregation and content of TXB(2) and 6-keto-PGF1α in plasma and the aortic wall. CONCLUSION: These results suggest that QSLC can be used in the prevention and treatment of thrombotic diseases, and that its mechanism of action may be related to inhibition of platelet aggregation.

Anti-HBV agents derived from botanical origin.

There are 350,000 hepatitis B virus (HBV) carriers all over the world. Chronic HBV infection is at a high risk of developing liver cirrhosis and hepatocelluar carcinoma (HCC), and heavily threatened people's health. Two kinds of drugs approved by FDA for anti-HBV therapy are immunomodulators (interferon α, pegylated-interferon α) and nucleos(t)ide analogues (lamivudine, adefovir dipivoxil, entecavir, telbivudine, and tenofovir disoproxil fumarate). These drugs have been proved to be far from being satisfactory due to their low specificity, side effects, and high rate of drug resistance. There is an urgent need to discover and develop novel effective anti-HBV drugs. With vast resources, various structures, diverse biological activities and action mechanisms, as well as abundant clinical experiences, botanical agents become a promising source of finding new anti-HBV drugs. This review summarizes the recent research and development of anti-HBV agents derived from botanical origin on their sources and active components, inhibitory effects and possible toxicities, as well as action targets and mechanisms, and also addresses the advantages and the existing shortcomings in the development of botanical inhibitors. This information may not only broaden the knowledge of anti-HBV therapy, and offer possible alternative or substitutive drugs for CHB patients, but also provides considerable information for developing new safe and effective anti-HBV drugs.

Inhibitory effects of Qushuanling Capsule () on thrombus formation and platelet aggregation in rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of Qushuanling Capsule ( QSLC) on thrombus formation and platelet aggregation in rats.</p><p><b>METHODS</b>Arteriovenous bypass, venous thrombosis, and middle cerebral artery thrombosis models were used in rats to investigate the anti-thrombotic effects of QSLC, a compound of nine Chinese herbs. The platelet aggregation induced by adenosine diphosphate (ADP), thrombin or arachidonic acid (AA), as well as the contents of thromboxane B(2) (TXB(2)) and 6-keto-prostaglandin F1α (6-keto-PGF1α) in rat plasma and aortic walls, were determined to investigate the possible mechanisms of the anti-thrombotic effects of QSLC.</p><p><b>RESULTS</b>After oral administration with QSLC for 7 days, arteriovenous bypass thrombosis was obviously suppressed compared with the model group, venous thrombosis was also obviously suppressed, rat behaviors were obviously improved, and brain infarct size as well as water content were also reduced. The platelet aggregation induced by ADP or thrombin was inhibited by QSLC, but the drug had no effect on AA-induced platelet aggregation and content of TXB(2) and 6-keto-PGF1α in plasma and the aortic wall.</p><p><b>CONCLUSION</b>These results suggest that QSLC can be used in the prevention and treatment of thrombotic diseases, and that its mechanism of action may be related to inhibition of platelet aggregation.</p>

Controlled observation of the efficacy between floating acupuncture at Tianying point and warm-needling therapy for supraspinous ligament injury / 中国针灸

<p><b>OBJECTIVE</b>To compare the efficacy difference in the treatment of supraspinous ligament injury between floating acupuncture at Tianying point and the conventional warm needling therapy.</p><p><b>METHODS</b>Ninety patients were randomized into a floating acupuncture group and a warm needling group, 45 cases in each one. In the floating acupuncture group, the floating needling technique was adopted at Tianying point. In the warm needling group, the conventional warm needling therapy was applied at Tianying point as the chief point in the prescription. The treatment was given 3 times a week and 6 treatments made one session. The visual analogue scale (VAS) was adopted for pain comparison before and after treatment of the patients in two groups and the efficacy in two groups were assessed.</p><p><b>RESULTS</b>The curative and remarkably effective rate was 81.8% (36/44) in the floating acupuncture group and the total effective rate was 95.5% (42/44), which were superior to 44.2% (19/43) and 79.1% (34/43) in the warm needling group separately (P < 0.01, P < 0.05). VAS score was lower as compared with that before treatment of the patients in two groups (both P < 0.01) and the score in the floating acupuncture group was lower than that in the warm needling group after treatment (P < 0.01). Thirty-six cases were cured and remarkably effective in the floating acupuncture group after treatment, in which 28 cases were cured and remarkably effective in 3 treatments, accounting for 77.8 (28/36), which was apparently higher than 26.3 (5/19) in the warm-needling group (P < 0.01).</p><p><b>CONCLUSION</b>The floating acupuncture at Tianying point achieves the quick and definite efficacy on supraspinous ligament injury and presents the apparent analgesic effect. The efficacy is superior to the conventional warm-needling therapy.</p>

Polygonum cuspidatum Extract Induces Anoikis in Hepatocarcinoma Cells Associated with Generation of Reactive Oxygen Species and Downregulation of Focal Adhesion Kinase.

Anoikis has been recognized as a potential target for anticancer therapy. Polygonum cuspidatum (Huzhang) is a frequently used Chinese herb in hepatocarcinoma. In present study, we evaluated the effects of Polygonum cuspidatum extract (PCE) in hepatocarcinoma cells in suspension. The results showed that PCE inhibited the proliferation of hepatocarcinoma cells in suspension in a dose- and time-dependent manner. PCE also inhibited anchorage-independent growth of hepatocarcinoma cells in soft agar. PCE induced anoikis in human hepatocarcinoma Bel-7402 cells accompanied by caspase-3 and caspase-9 activation and poly(ADP-ribose) polymerase cleavage, which was completely abrogated by a pan caspase inhibitor, Z-VAD-FMK. In addition, PCE treatment induced intracellular reactive oxygen species (ROS) production in Bel-7402 cells. NAC, an ROS scavenger, partially attenuated PCE-induced anoikis and activation of caspase-3 and caspase-9. Furthermore, PCE inhibited expression of focal adhesion kinase (FAK) in Bel-7402 cells. Overexpression of FAK partially abrogated PCE-induced anoikis. These data suggest that PCE may inhibit suspension growth and induce caspase-mediated anoikis in hepatocarcinoma cells and may relate to ROS generation and FAK downregulation. The present study provides new insight into the application of Chinese herb for hepatocarcinoma treatment.

Liver Yin deficiency tonifying herbal extract induces apoptosis and cell senescence in Bel-7402 human hepatocarcinoma cells.

Liver cancer ranks as the fifth most prevalent malignancy of all cancers worldwide. According to the principles of traditional Chinese medicine, liver Yin deficiency is a common clinical syndrome of liver cancer, and tonifying liver Yin is a common treatment method for liver cancer. However, no hepatocarcinoma-specific liver Yin tonifying formula has yet been established. In the present study, we established a liver cancer-specific combination of herbs, which we term liver Yin tonifying formula (LYTF). We found that LYTF inhibits the proliferation of Bel-7402 cells in a dose- and time-dependent manner. LYTF induces apoptosis in Bel-7402 cells, which is accompanied by activation of caspases-8, -9 and -3. Pan-caspase blocking completely abrogates LYTF-induced apoptosis and partially abrogates LYTF-induced proliferation inhibition. LYTF also induces cell senescence, as indicated by a large and flattened morphology, senescence-activated ß-galactosidase-positive staining and G0/G1 cell cycle arrest, accompanied by the up-regulation of p16 and p21 and the down-regulation of retinoblastoma protein phosphorylation. These findings suggest that LYTF is effective in inhibiting the growth and survival of hepatocarcinoma cells through the induction of apoptosis and cell senescence. Our study also provides insight into traditional Chinese medicine methods used for the treatment of liver cancer.

[Effects of resveratrol on apoptosis and ROS production in Hepa 1-6 hepatocarcinoma cells].

OBJECTIVE: To observe the effects of resveratrol on apoptosis and ROS production in murine hepatocarcinoma Hepa 1-6 cells in vitro. METHODS: Hepa 1-6 cells were treated with different dose of resveratrol (20 micromol/L, 40 micromol/L, 80 micromol/L). Cell proliferation was detected with MTT assay at 24 h, 48 h and 72 h. Hoechst 33258 staining was used to visualize apoptotic morphology. Cell apoptosis was detected by flow cytometry analysis. Activated caspase-3 was detected by western blot. Intracellular ROS production was observed by 2',7'-dichlorofluorescin diacetate (DCF-DA) staining. RESULTS: Compared with the control, upon treatment with 20-80 micromol/L resveratrol for 24 h, 48 h or 72 h, the proliferation of Hepa 1-6 cells was significantly inhibited in a time-dose dependent manner. 20-80 micromol/L resveratrol also induced apoptosis and apoptotic morphology change in Hepa 1-6 cells accompanied with caspase-3 activation and ROS generation. CONCLUSION: Resveratrol could inhibit cell proliferation, induce apoptosis in murine hepatocarcinoma Hepa 1-6 cells, and the mechanism may associated with caspase-3 activation and ROS production.