The Antitumour Effect of Prunella vulgaris Extract on Thyroid Cancer Cells In Vitro and In Vivo.

Prunella vulgaris, a traditional Chinese medicine, has been used to treat various benign and malignant tumours for centuries in China. In our previous studies, Prunella vulgaris extract (PVE) was shown to promote apoptosis in differentiated thyroid cancer (DTC) cells. However, whether other mechanisms are involved in the antitumour effect of PVE in thyroid cancer (TC) cells remains unclear. The present study aimed to investigate the antiproliferative and antimigratory effects of PVE on TC cell lines both in vitro and in vivo. First, the TPC-1 and SW579 human TC cell lines were screened by MTT assay for their high level of sensitivity to PVE. Then, the results of cell growth curve and colony formation assay and cell cycle analyses, wound healing, and migration assays demonstrated that PVE inhibited the proliferation and migration of TPC-1 and SW579 cells. Moreover, the antitumour effect of PVE was verified in a subcutaneous xenotransplanted tumour model. Next, MKI67, PCNA, CTNNB1, and CDH1 were screened by qRT-PCR for their significantly differential expression levels in xenograft tissue with and without PVE treatment, and expression of MKI67, PCNA, and CDH1 was verified by Western blot. Finally, an integrated bioinformatics analysis containing protein-protein interaction network, KEGG pathway, and GO analysis was conducted to explore more potential antitumour mechanisms of PVE. In summary, PVE could inhibit the proliferation and migration of TC cells both in vitro and in vivo, which may have been achieved by modulation of the expression of MKI67, PCNA, and CDH1. These data suggest that PVE has the potential to be developed into a new anticancer drug for the treatment of TC.

Synthesis and biological evaluation of a novel ß-D-2'-deoxy-2'-α-fluoro-2'-ß-C-(fluoromethyl)uridine phosphoramidate prodrug for the treatment of hepatitis C virus infection.

A novel ß-D-2'-deoxy-2'-α-fluoro-2'-ß-C-(fluoromethyl)uridine phosphoramidate prodrug (1) has been synthesized. This compound exhibits submicromolar-level antiviral activity in vitro against HCV genotypes 1b, 1a, 2a, and S282T replicons (EC50 = 0.18-1.13 µM) with low cytotoxicity (CC50 > 1000 µM). Administered orally, prodrug 1 is well tolerated at doses of up to 4 g/kg in mice, and produces a high level of the corresponding triphosphate in rat liver.

Protective effects of 20(s)-protopanaxtriol on viral myocarditis infected by coxsackievirus B3.

OBJECTIVE: Coxsackievirus B3 (CVB3) is a dominant causative agent for viral myocarditis. So far, effective therapies for the treatment of the disease are not available. 20(S)-Protopanaxtriol is a major component of Panax pseudoginseng and has been clinically used for the treatment of heart diseases. However, it is not known whether 20(S)-protopanaxtriol exerts any anti-viral effects. Thus, the aim of this study was to investigate the therapeutic effects of 20(S)-protopanaxtriol against CVB3 in vivo and in vitro. METHODS: The antiviral effects of 20(S)-protopanaxtriol in vitro were evaluated in HeLa cells infected by CVB3. Then, we examined the protective effects of 20(S)-protopanaxtriol on CVB3-induced myocarditis in BALB/c mice. These mice were treated with 20(S)-protopanaxtriol at doses of 100-400 mg·kg(-1)·day(-1) for 7 days and compared with the controls. RESULTS: We found that 20(S)-protopanaxtriol possessed potent antiviral effects on CVB3 in vitro. Compared with control mice, virus titers and pathological changes in the hearts were significantly decreased in the 20(S)-protopanaxtriol-treated group. Furthermore, biochemical markers of myocardial injury such as plasma lactate dehydrogenase and creatine kinase were decreased to normal levels. CONCLUSIONS: These data provide the possibility that 20(S)-protopanaxtriol can be used as a potential therapeutic means for treatment of viral myocarditis.

Phyllaemblicin B inhibits Coxsackie virus B3 induced apoptosis and myocarditis.

Coxsackie virus B3 (CVB3) is believed to be a major contributor to viral myocarditis since virus-associated apoptosis plays a role in the pathogenesis of experimental myocarditis. In this study, we investigated the in vitro and in vivo antiviral activities of Phyllaemblicin B, the main ellagitannin compound isolated from Phyllanthus emblica, a Chinese herb medicine, against CVB3. Herein we report that Phyllaemblicin B inhibited CVB3-mediated cytopathic effects on HeLa cells with an IC(50) value of 7.75+/-0.15microg/mL. In an in vivo assay, treatment with 12mgkg(-1)d(-1) Phyllaemblicin B reduced cardiac CVB3 titers, decreased the activities of LDH and CK in murine serum, and alleviated pathological damages of cardiac muscle in myocarditic mice. Moreover, Phyllaemblicin B clearly inhibited CVB3-associated apoptosis effects both in vitro and in vivo. These results show that Phyllaemblicin B exerts significant antiviral activities against CVB3. Therefore, Phyllaemblicin B may represent a potential therapeutic agent for viral myocarditis.

[The experimental research about the effect of Prunella vulgaris L. on Raji cells growth and expression of apoptosis related protein].

OBJECTIVE: To investigate the anti-lymphoma effect of Prunella vulgaris L. in order to offer exprimental data for the treatment of lymphoma with Prunella vulgaris L. in clinic. METHODS: Effect of Prunella vulgaris L. injection on inhibition ratio of cell growth of Raji cells and IC50 were tested by MTT assay. The growth curve line of Raji cells was drawn also by MTT assay. The cellular morphology was observed by invert microscope, Giemas staining and MTT assay. The expression of apoptosis related protein bcl-2, bax was measured by immunocytochemistry and the quantitative analysis was made with figure analysis system. RESULTS: 1. Prunella vulgaris L. could obviously suppress the cell proliferation of Raji cells in a concentration-dependent manner (r = 0.97). The IC50 was 0.118 mg/ml. 2. After Raji cells was reacted with injection of Prunella vulgaris L. (50 mg/ml) , the morphlogical of apoptosis were observed by invert microscope, Giemsa staining and MTT assay. RESULTS: The results of immunocytochemistry showed that after Raji cells were treated by the injection of Prunella vulgaris L. (50 mg/ml) for 48 hours, the expression of bcl-1 was up-regulated, and the expression of bax was down-regulated. The differences between process group and control group were significant (P < 0.01). CONCLUSION: Prunella vulgaris L. can suppress the proliferation of Raji cells and may be a new anti-lymphoma drug. Inducing the apoptosis of Raji cells maybe one of anti-lymphoma mechanisms.

[Reversal of multidrug resistance in drug-resistant cell line EAC/ADR by cepharanthine hydrochloride and its mechanism].

AIM: To investigate the correlation between reversal effect of cepharanthine hydrochloride (CH) on multidrug resistance (MDR) in drug-resistant cell line EAC/ADR and the nuclear transcription factor-KB (NF-KB). METHODS: Cytotoxicity was determined by the tetrazolium (MTT) assay in vitro. An EAC/ADR cell homograft model was established to investigate the effect of CH on reversing MDR in vivo. The constitutive activity and activation of NF-KB by drugs were measured by Dot-Enzyme-linked Immune Sorbent Assay (Dot-ELISA). RESULTS: CH was shown to potentiate the cytotoxicity of ADR, a 13- fold reversal effect of resistance was achieved in vitro. In mice bearing EAC/ADR cell homografts, CH was found to prolong the survival time of animals bearing tumor. Increase in life span over control was 75. 37%. In addition, the constitutive activity of NF-KB and activation of NF-KB by chemotherapy were lowered by CH. CONCLUSION: The findings suggest that CH is able to reverse drug resistance and its mechanism may be related to suppressing the constitutive activity and activation of NF-KB by drugs.

[Regulation of telomerase activity and cell cycle of K562 cells by oridonin].

AIM: To investigate the effect of oridonin (ORI) on telomerase activity and cell cycle of human leukemic cell line K562 cells. METHODS: Immunohistochemistry (IHC) technique was used to determine the expression of hTERT or C-myc. Telomerase activity was detected with TRAP-PCR-ELISA assay. In addition, the percentages of K562 cells in different cell cycle were determined by flow cytometry (FCM) at 24th and 48th hours separately after adding the different concentrations of ORI. RESULTS: After the K562 cells were treated with ORI at 3.43 micromol x L(-1) for 48 h, the expression of hTERT and C-myc decreased obviously. There was statistical significant (P < 0.05) difference between experimental groups and the normal controls. In addition, the telomerase activity of K562 cells was significantly inhibited by ORI at the dose of 3.43 micromol x L(-1) for 48 h. At the same time, the cell cycle distribution changed, the percentage of G0/G1 or G2/M stages cells increased and that of the S stage cells decreased after ORI was added. CONCLUSION: ORI can effectively inhibit telomerase activity in K562 cells. Arresting cell cycle and decreasing the expression of hTERT and C-myc may be the mechanism of action.