Erratum: Jiedu Sangen Decoction Reverses Epithelial-to-mesenchymal Transition and Inhibits Invasion and Metastasis of Colon Cancer via AKT/GSK-3ß Signaling Pathway: Erratum.

[This corrects the article DOI: 10.7150/jca.32873.].

Jiedu Sangen decoction inhibits chemoresistance to 5-fluorouracil of colorectal cancer cells by suppressing glycolysis via PI3K/AKT/HIF-1α signaling pathway.

Drug resistance is a major obstacle in the development of effective colorectal cancer (CRC) therapy. Our study aimed to explore the reversal abilities of Jiedu Sangen decoction (JSD) on the 5-fluorouracil (5-FU) resistance and its underlying molecular mechanisms. Expression changes in HIF-1 of CRC tissues were firstly revealed by bioinformatics analysis. Afterwards, cell viabilities of JSD and 5-FU treatments on 5-FU resistant human colon cancer cells (HCT-8/5-FU) were determined. Expressions of phosphoinositide 3-kinase (PI3K), protein kinase B (AKT)/p-AKT, hypoxia-inducible factor 1 (HIF-1α), as well as glycolysis related proteins such as L-lactate dehydrogenase A (LDHA), Glucose transporter type 1 (Glut1), Hexokinase 2 (HKII), and cysteinyl aspartate specific proteinase (Caspase) family members in HCT-8/5-FU cells, HIF-1α silenced HCT-8/5-FU cells and tumor tissues were detected by western blotting. HIF-1α was found over expressed in CRC tissues according to public available datasets in Oncomine. Growth inhibition rates of HCT-8/5-FU cells were increased along with the increase of JSD concentrations. JSD caused down-regulated HIF-1α, PI3K, AKT/p-AKT, HKII and Glut1, as well as up-regulated Caspase3 and Caspase9 in HCT-8/5-FU cells and tumor tissues. In HIF-1α silenced HCT-8/5-FU cells, synergistic group showed significantly reduced expression levels of PI3K, AKT, p-AKT. Additionally, up-regulated expressions of Caspase6 and Caspase7 were observed. JSD combined with 5-FU also exhibited obvious inhibitory efficiency on tumor growth in vivo. JSD may reverse 5-FU resistance by suppressing glycolysis via PI3K/AKT/HIF-1α signaling pathway, thereby inhibiting glycolysis and induce apoptosis to enhance anti-tumor activity.

Jiedu Sangen Decoction Reverses Epithelial-to-mesenchymal Transition and Inhibits Invasion and Metastasis of Colon Cancer via AKT/GSK-3ß Signaling Pathway.

Ethnopharmacology relevance: Jiedu Sangen Decoction (JSD), an empirical prescription of Traditional Chinese Medicine (TCM), has been reported to inhibit invasion and metastasis of colon cancer in our previous study. The aim of this study was to investigate the mechanism of JSD-triggered inhibition of invasion and metastasis in colon cancer. Methods: In vitro, AKT1 knockdown (si-AKT1) or overexpression (oe-AKT1) cells were successfully constructed both in SW480 and SW620 cell lines. Si-AKT1 and oe-AKT1 cells were then treated with or without JSD. Cell invasion, metastasis potential and expression of epithelial-mesenchymal transformation (EMT)-related and AKT1/GSK-3ß proteins were then observed by wound healing, transwell, and western blot assays. In vivo, liver metastasis model mice were developed by inoculating SW480 cells. After JSD diet intervention, living fluorescence imaging and weight measurements were carried out to investigate JSD induced inhibition effects on liver metastasis of colon cancer. Immunohistochemistry and western blot assays were performed to observe tissue features and detect protein expression. Results: Invasion and metastasis potential, as well as EMT of colon cancer, can be markedly inhibited by JSD treatment or AKT1 knockdown, while enhanced by AKT1 overexpression. JSD-induced inhibition effects were significantly weakened when AKT1 was knocked down, while clearly enhanced when AKT1 was overexpressed. Additionally, JSD could lead to an increase in expression of E-cadherin, and a decrease in expression of N-cadherin, Vimentin, p-AKT1, AKT1, p- GSK-3ß, Snail, Slug, and Twist in colon cancer cells. Conclusion: JSD reverses EMT and inhibits invasion and metastasis of colon cancer through the AKT/GSK-3ß signaling pathway.

[Beta-HIVS combined cisplatin inhibited activities of human ovarian cancer cell line SKOV3 in vitro].

OBJECTIVE: To study the effect of beta-hydroxyisovaleryl shikonin (beta-HIVS) combined cisplatin on activities of ovarian cancer cell line SKOV3 in vivo and its possible mechanisms. METHODS: Cells were divided into the blank control group and six beta-HIVS groups (2 - 30 micromol/L). Effect of beta-HIVS at different concentrations on the activities of ovarian cancer cell line SKOV3 was detected using MTT assay. SKOV3 cells were treated with cisplatin (10, 20, and 40 micromol/L) and beta-HIVS (0.25, 1, and 2.5 micromol/L) combined cisplatin. Effect of beta-HIVS combined cisplatin on the activities of ovarian cancer cell line SKOV3 was determined by MTT assay. The expression of Bcl-2 and Bax after treated by different concentrations of beta-HIVS was detected by Western blot. RESULTS: The activities of SKOV3 were inhibited by different concentrations of beta-HIVS dose-dependently. The 50% inhibition rate (IC50) was 7.37 micromol/L. There was statistical difference in IC50 between each concentration beta-HIVS group and the blank control group (P < 0.05). There was statistical difference in IC50 between the beta-HIVS (1 and 2.5 micromol/L) combined cisplatin groups and the cisplatin group (P < 0.05, P < 0.01). The synergistic effect on beta-HIVS showed dose-dependent manner. Results of Western blot showed beta-HIVS at different concentrations (5, 7.5, and 10 micromol/L) could obviously up-regulate the expression level of Bax protein and inhibit the expression level of Bcl-2 protein, showing statistical difference when compared with the control group (P < 0.01). CONCLUSIONS; HIVS could obviously inhibit in vitro growth of SKOV3 in a dose-dependent manner. With the range of concentration, beta-HIVS showed synergetic effect with cisplatin. Besides, along with increasing beta-HIVS concentrations, the synergetic effect was more significant. The synergetic effect might accelerate the apoptosis of SKOV3 through up-regulating Bax expression and inhibiting Bcl-2 expression.

[Surgical resection versus radiofrequency ablation for primary hepatocellular carcinoma of 3-5 cm in diameter: a meta-analysis].

OBJECTIVE: To compare the therapeutic effects of surgical hepatic resection (HR) and radiofrequency ablation (RFA) in treatment of primary hepatocellular carcinoma of 3-5 cm in diameter. METHODS: The databases PubMed, CBMdisc, CNKI, WanFang Data and VIP databases were searched for controlled clinical trials on evaluating the efficacy between RFA and HR in treatment of primary hepatocellular carcinoma of 3-5 cm in diameter published from January 1990 to February 2014. Two reviewers independently screened the literature, extracted the data and assessed the methodological quality of the studies included. Then the meta-analysis was performed by using RevMan5.0 software. RESULTS: Eleven controlled clinical trials were included, including one randomized controlled trial and 10 non-randomized controlled trials. A total of 811 patients were involved: 404 patients were treated with RFA as the initial treatment and 407 patients with surgical resection. Meta-analysis showed that for a single lesion with diameter of 3-5 cm of primary hepatocellular carcinoma, the 3-， 5-year disease-free survival rates in HR group was significantly higher than those in RFA group (all P<0.05). There were no significant difference in the 1-， 3-， 5-year overall survival rates and 1-year disease-free survival rate between RFA group and HR group (P>0.05). For 1-2 nodules with diameters of 3-5 cm of primary hepatocellular carcinoma, the 3-， 5-year disease-free survival rates and 5-year overall survival rates in HR group was significantly higher than those in RFA group (all P<0.05). No significant difference in 1-， 3-year overall survival rates and 1-year disease-free survival rate was found between RFA group and HR group (P>0.05). For maximum nodule of 3-5 cm of multiple primary hepatocellular carcinoma, the 5-year overall survival rates in HR group was significantly higher than that in RFA group (all P<0.05). No significant difference in 1-， 5-year overall survival rates was noted between RFA group and HR group (P>0.05). CONCLUSION: For primary hepatocellular carcinoma of 3-5 cm in diameter, HR is better than RFA. For the limitation of quality and quantity of included studies, this conclusion needs to be confirmed by more high quality studies.

[Jiedu sangen decoction intervened carcinoma-associated fibroblasts and inhibited migration and invasion of colon cancer: an experimental research].

OBJECTIVE: To observe the effect of Jiedu Sangen Decoction (JSD, consisting of Polygonum cuspidatum, Geum Japonicum Thumnb, Radix Actinidiae Chinensis) on the migration capability of colon cancer CT-26 cells were observed, and on expressions of carcinoma-associated fibroblasts (CAFs) such as transforming growth factor-beta1 (TGF-beta1), matrix metalloproteinase 9 (MMP-9), and alpha-smooth muscle actin (alpha-SMA). METHODS: The BALB/C mice were subcutaneously inoculated with colon cancer CT-26 cells (1.2 x 10(6)mL) and then randomly divided into 3 groups, i.e., the normal control group, the model group, the JSD treated group. The effects of three different serums on the migration ability of colon cancer CT-26 cells were observed using Transwell. The expression quantities of TGF-beta1 and MMP-9 in the supernatant of CAFs were detected using ELISA. The mRNA expression quantities of TGF-beta1 and alpha-SMA in CAFs were detected by real-time fluorescence quantitative PCR. RESULTS: The number of semi-permeable film cells in the JSD treated group significantly decreased, when compared with the model group, showing statistical significance (P < 0.01). Compared with the model group, the expressions of TGF-beta1 and MMP-9 in the supernatant of CAFs decreased in the JSD treated group at 24 and 48 h, showing statistical difference (P < 0.05, P < 0.01). Compared with the model group, the mRNA expressions of TGF-beta1 and alpha-SMA in the JSD treated group obviously decreased, showing statistical difference (P < 0.01). CONCLUSION: JSD could decrease expressions of TGF-beta1 and MMP-9 in the supernatant of CAFs, lower mRNA expressions of alpha-SMA and TGF-beta1, which might be possible mechanisms for inhibiting the migration and invasion of tumor cells.

Inhibition of breast cancer metastasis via PITPNM3 by pachymic acid.

Breast cancer metastasis is the most common cause of cancer-related death in women. Thus, seeking targets of breast tumor cells is an attractive goal towards improving clinical treatment. The present study showed that CCL18 from tumor-associated macrophages could promote breast cancer metastasis via PITPNM3. In addition, we found that pachymic acid (PA) could dose-dependently inhibit migration and invasion of MDA-MB-231 cells, with or without rCCL18 stimulation. Furthermore, evidence was obtained that PA could suppress the phosphorylation of PITPNM3 and the combination of CCL18 and PITPNM3. Therefore, we speculate that PA could inhibit breast cancer metastasis via PITPNM3.

[Effect of shuizhongcao granule on cellular immune function of experimental animal with recurrent aphthous stomatitis].

OBJECTIVE: To investigate the effect of Shuizhongcao Granule (SZCG) on cellular immune function in animal model of recurrent aphthous stomatitis (RAS). METHODS: Experimental animal model of RAS were established by immunological method. The modeled animals were randomized into 4 groups, Group A was the model control group, Group B was treated by Levamisole (2.5 mg/mL), Group C and D was treated with high-dose (0.5 g/mL) and low-dose (0.24 g/mL) SZCG respectively. The medication was administered via gastric perfusion, starting from the 7th week of the experiment, and continued for 28 days. Meanwhile, a normal control group was set up. Percentage of T-lymphocyte subsets, CD3, CD4 and CD8, and serum levels of interleukin 10 and 12 (IL-10 and IL-12) in peripheral blood were detected before and after treatment. RESULTS: Percentages of CD3, CD4 and CD4/CD8 ratio in the model control group were 40.50 +/- 7.46%, 30.80 +/- 5.33% and 79.56 +/- 8.32 respectively, all lower than those in the normal control group (P<0.05); level of serum IL-10 was higher (8.02 +/- 0.53 ng/L) and that of IL-12 was lower (15.51 +/- 1.35 ng/L) in the model animal than those in normal control (P<0.05). After treatment, levels of CD3, CD4, CD4/CD8 and IL-12 in Group B were 55.30 +/- 6.33%, 43.50 +/- 5.33%, 99.29 +/- 13.84 and 17.72 +/- 1.70 ng/L, those in Group C were 61.50 +/- 4.32%, 47.20 +/- 4.37%, 103.30 +/- 7.42 and 18.18 +/- 1.54 ng/L, and in Group D were 58.50 +/- 6.03%, 42.80 +/- 3.17%, 110.27 +/- 12.85 and 17.74 +/- 1.96 ng/L, respectively, all were higher than those in Group A, but levels of IL-10 in the three treatment groups were lower than that in Goup A (P<0.05 or P<0.01). CONCLUSION: The immune function of RAS model animals is in the suppressed condition; SZCG can improve the immune suppression to achieve its therapeutic effect on the disease.

[Effect of guilu erxianjiao in suppressing splenic T-lymphocyte apoptosis in mice undergoing chemotherpy].

OBJECTIVE: To investigate the effect of Guilu Erxianjiao (GLEXJ) in suppressing chemotherapy induced cell apoptosis in mice and to discuss its possible acting mechanism. METHODS: Seventy-two tumor-bearing Kunming mice were randomly divided into 6 groups, Group A, treated with normal saline; Group B treated with cyclophosphamide (CTX), Group C treated with Chinese herbal medicine, and Group D-F, the three combined treated group received CTX plus high, moderate and low dose GLEXJ. All mice were sacrificed after 9-day medication, their splenic tissues were taken for determining T-lymphocyte apoptosis with flow cytometer, the expression of bcl-2 mRNA and Caspase-3mRNA detected by RT-PCR. RESULTS: As compared with Group A, FITC+/PI- cell proportion of spleen T-lymphocyte cells and Caspase-3mRNA expression were higher, while bcl-2 mRNA expression was lower in Group B; as compared with Group B, the former two indexes were lower and the latter was higher in the three combined treated groups (all P <0.05). CONCLUSION: GLEXJ could efficiently suppress the splenic T-lymphocyte apoptosis in tumor bearing mice undergoing chemotherapy, one of its mechanisms may be through up-regulating of bcl-2 mRNA expression and down-regulating of Caspase-3mRNA expression.