Astragaloside IV and Saponins of Rhizoma Polygonati Cure Cyclophosphamide-Induced Myelosuppression in Lung Adenocarcinoma via Down-Regulating miR-142-3p.

Our previous study revealed that Shuanghuang Shengbai granule could cure the myelosuppression induced by cyclophosphamide (CTX) in lung cancer. However, its hematopoietic effects and molecular mechanisms remain not fully understood. Therefore, this study was intended to investigate the effects and the underlying mechanisms of Astragaloside IV (AS) and saponins of rhizoma polygonati (SRP), the two main bioactive ingredients of Shuanghuang Shengbai granule, on CTX-induced myelosuppression. CTX inhibited the proliferation and promoted apoptosis in bone marrow hematopoietic stem cells (BMHSCs), accompanied by the increased expression of miR-142-3p. AS and/or SRP treatment could alleviate CTX-induced cell injury and suppress the expression of miR-142-3p. Over-expression of miR-142-3p partially reversed the therapeutic effect of AS and/or SRP on CTX-induced cell injury in BMHSCs. Further mechanism exploration discovered that HMGB1 was the target gene of miR-142-3p, and miR-142-3p negatively regulated the expression of HMGB1. To further explore the function of AS and/or SRP in vivo, we constructed a lung cancer xenograft combined with CTX-induced myelosuppression mouse model, and we found that AS and SRP remarkably reversed the CTX-induced reduction of white blood cells, bone marrow nucleated cells, and thymus index in vivo and did not affect the chemotherapy effect of lung cancer. Collectively, our results strongly suggested that AS and SRP could improve the hematopoietic function of myelosuppressed lung cancer mice, and their effects may be related to the inhibition of miR-142-3p expression in BMHSCs.

Ursolic acid induces apoptosis and anoikis in colorectal carcinoma RKO cells.

BACKGROUND: Ursolic acid (UA) is an anti-cancer herbal compound. In the present study, we observed the effects of UA on anchorage-dependent and -independent growth of human colorectal cancer (CRC) RKO cells. METHODS: RKO cells were cultured in conventional and detached condition and treated with UA. Cell viability was evaluated by CCK-8 assay. Cell cycle was analyzed by flow cytometry. Apoptosis was identified by Hoechst 33258 staining and flow cytometry analysis. Activities of caspases were measured by commercial kits. Reactive oxygen species (ROS) was recognized by DCFH-DA fluorescent staining. Anoikis was identified by EthD-1 fluorescent staining and flow cytometry analysis. Expression and phosphorylation of proteins were analyzed by western blot. RESULTS: UA inhibited RKO cell viability in both a dose- and time-dependent manner. UA arrested the cell cycle at the G0/G1 phase, and induced caspase-dependent apoptosis. UA inhibited Bcl-2 expression and increased Bax expression. In addition, UA up-regulated the level of ROS that contributed to UA activated caspase-3, - 8 and - 9, and induced apoptosis. Furthermore, UA inhibited cell growth in a detached condition and induced anoikis in RKO cells that was accompanied by dampened phosphorylation of FAK, PI3K and AKT. UA also inhibited epithelial-mesenchymal transition (EMT) as indicated by the down-regulation of N-Cad expression and up-regulation of E-Cad expression. CONCLUSIONS: UA induced caspase-dependent apoptosis, and FAK/PI3K/AKT singling and EMT related anoikis in RKO cells. UA was an effective anti-cancer compound against both anchorage-dependent and -independent growth of RKO cells.

Ursolic acid potentiated oxaliplatin to induce apoptosis in colorectal cancer RKO cells.

Ursolic acid (UA) is found in multiple anticancer herbs and has shown anticancer effects in colorectal cancer (CRC) cells. The present study aimed to observe the effects of a combination of UA and oxaliplatin (Oxa), a frequently used chemotherapeutic drug in CRC, on human CRC RKO cells. The results showed that UA and Oxa synergistically inhibited the proliferation of RKO cells. A combination of UA and Oxa induced apoptosis in RKO cells and increased the activities of caspase-3, caspase-8, and caspase-9. Z-VAD-FMK, a caspase inhibitor, significantly antagonized UA- and Oxa-activated caspase-3, caspase-8, and caspase-9 and induced apoptosis. In addition, UA and Oxa downregulated the expression of X-linked inhibitor of apoptosis (XIAP) and Survivin in RKO cells. These observations suggested that a combination of UA and Oxa elicited synergistically anticancer effects in RKO cells and provided new evidence for potential application of UA and Oxa for CRC treatment.

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.

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.

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.

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.

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.

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.

[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.

Inflammation, macrophage in cancer progression and chinese herbal treatment.

Inflammation is associated with cancer development, and has been recognized as the seventh hallmarks of the cancer. Cancer-related inflammation can be activated by genetic or epigenetic changes in cancer cells (intrinsic pathway) or mediated by tumor-infiltrating immune cells (extrinsic pathway). Immune cells involved in cancer-related inflammation mainly including tumor-associated macrophages or M2 macrophages, neutrophils, dendritic cells, mast cells, and lymphocytes. As major players of the cancer-related inflammation, M2 macrophages, secreting various of growth factors, immunomodulatory cytokines and matrix metalloproteinases, participate in remodeling of extracellular matrix, contribute to cancer invasion and metastasis, angiogenesis, and inhibit anti-cancer immunity. Inflammation has been considered as an important target for cancer therapy. Some Chinese herbal ingredients have been confirmed to be effective in inhibit inflammation related gene expression in cancer cells, such as COX-2 and NF-B. However, there is a shortage of study on Chinese herb or herbal ingredient against extrinsic cancer inflammation, especially in tumor-associated macrophages. Related studies may provide new insight into cancer treatment.

Modified Yi Guan Jian, a Chinese herbal formula, induces anoikis in Bel-7402 human hepatocarcinoma cells in vitro.

Liver cancer is the fifth most common malignancy worldwide. Liver YIN deficiency is a common clinical syndrome of traditional Chinese medicine in liver cancer. Yi Guan Jian is an ancient classic liver YIN tonifying herbal formula used for the treatment of liver disease with liver YIN deficiency, which is also currently used for liver cancer treatment. However, as an ancient formula, Yi Guan Jian (YGJ) is not entirely suitable for liver cancer treatment. In the present study, we optimized the prescription of YGJ according to the current principles of Chinese herbal medication, and evaluated the anticancer effects of modified Yi Guan Jian (MYGJ) in Bel-7402 human hepatocarcinoma cells. The results show that MYGJ inhibited the growth of Bel-7402 cells in adherent or in suspension cultures, and was more effective in Bel-7402 cells in suspension. MYGJ also inhibited anchorage-independent growth of Bel-7402 cells in soft agar. MYGJ induced anoikis in Bel-7402 cells accompanied by caspase-3, -8 and -9 activation, which was blocked by the pan-caspase inhibitor, Z-VAD-FMK. Furthermore, MYGJ inhibited the expression and phosphorylation of p38 MAPK in Bel-7402 cells. These findings suggest that MYGJ is sufficient to induce caspase-mediated anoikis in Bel-7402 cells in vitro, and may be associated with down-regulation of p38 MAPK. The present study also provides insight into the application of ancient Chinese herbal formulas.

Aqueous extract of Curcuma aromatica induces apoptosis and G2/M arrest in human colon carcinoma LS-174-T cells independent of p53.

Curcuma aromatica is a common Chinese herb for treating diseases with blood stasis and has been regarded as an anticancer herb in modern clinical practice. However, the anticancer effects and related molecular mechanisms of Curcuma aromatica remain unclear. In the present study, human colon carcinoma LS-174-T cell line with wild-type p53 was used as a model cell to evaluate the anticancer effects of aqueous extract of Curcuma aromatica (AECA). AECA inhibits LS-174-T cell proliferation in a dose- and time-dependent manner and colony formation in a dose-dependent manner. AECA treatment induces apoptosis accompanied by caspase-8, -9, and -3 activation in LS-174-T cells. Moreover, blocking the activities of these caspases with a specific inhibitor significantly protected LS-174-T cells from AECA-induced apoptosis. AECA treatment also induces G2/M phase arrest in LS-174-T cells. Expression of p53 was unchanged after AECA treatment; specific silence of p53 did not influence AECA-induced apoptosis and G2/M phase arrest. Further, the expression of cyclin B1 and CDK1 was reduced by AECA. This study suggests that AECA might be effective as an antiproliferative herb for colon carcinoma, the antitumor activity of AECA may involve both extrinsic and intrinsic apoptosis, and AECA induces G2/M phase arrest via downregulation of cyclin B1 and CDK1 and without the participation of p53.

[Senescence-inducing effects of Chinese herbal medicine Tenglong Buzhong Decoction on human colon carcinoma LS-174-T cells and the mechanism].

OBJECTIVE: Cell senescence is an important anti-cancer mechanism and may contribute to cancer therapeutic outcome. The present study observed the effects of Tenglong Buzhong Decoction (TLBZD), a Chinese herbal formula, on senescence in human colon carcinoma LS-174-T cells. METHODS: LS-174-T cells were treated with TLBZD, and morphology change was observed under a microscope. Cell senescence was identified by senescence-associated ß-galactosidase (SA-ß-gal) staining, and cell cycle was analyzed by flow cytometry. Expressions of p53, p21(WAF1/CIP1), p16 and RB and RB phosphorylation were detected by Western blot. RESULTS: After being treated with TLBZD, LS-174-T cells became enlarged and flattened by morphology; SA-ß-gal staining was positive and cell cycle was arrested in G0/G1. In addition, up-regulations of p21(WAF1/CIP1) and p16, as well as inhibition of RB phosphorylation were detected in response to TLBZD treatment. Expressions of p53 and RB were unchanged after TLBZD treatment. CONCLUSION: TLBZD is effective in inducing cell senescence in human colon carcinoma LS-174-T cells, which may relate to up-regulations of p21(WAF1/CIP1) and p16 and inhibition of RB phosphorylation.

[Effects of Tenglong Buzhong Decoction on proliferation and apoptosis of human colon carcinoma cell line LS174T].

OBJECTIVE: To observe the effects of Tenglong Buzhong Decoction (TLBZD), a compound traditional Chinese herbal medicine, on proliferation and apoptosis of colon carcinoma cell line LS174T in vitro. METHODS: Human colon carcinoma cell line LS174T and human colon epithelial cell line CRL-1790 were treated with different doses of TLBZD. Cell proliferation was detected with cell counting kit-8 (CCK-8) assay and clone formation assay. Cell cycle and apoptosis were detected by flow cytometry, and caspase-3, -8 and -9 activities in LS174T cells were detected by colorimetric assay. RESULTS: TLBZD had no obvious cytotoxicity in normal CRL-1790 cells. After 72-hour treatment of 1 mg/mL TLBZD, or 48- and 72-hour of 2 mg/mL TLBZD, or 24-, 48- and 72-hour of 5-20 mg/mL TLBZD, proliferation of LS174T cells was significantly inhibited. Clone formation of LS174T cells was significantly inhibited by 1 to 20 mg/mL TLBZD treatment. TLBZD at doses of 5 to 20 mg/mL also induced apoptosis and cell cycle arrest at G(0)/G(1) phase in LS174T cells. In addition, caspase-3, -8 and -9 activities were significantly elevated after 5 to 20 mg/mL TLBZD treatment. CONCLUSION: TLBZD can inhibit cell proliferation, arrest cell cycle at G(0)/G(1) phase, and induce apoptosis in LS174T cells, which may be related to activating of caspase-3, -8 and -9.

[Clinical study on post-operative metastasis prevention of progressive stage of gastric cancer by weichang'an].

OBJECTIVE: To observe the effect of Weichang'an (WCA, a Chinese preparation) in preventing post-operative metastasis of progressive stage of gastric cancer. METHODS: A prospective randomized, controlled study was conducted by dividing the 148 patients of progressive staged gastric cancer after radical operation into the WCA group, the chemotherapy (CT) group and the WCA + CT group, to observe the survival rate, metastasis rate, quality of life (QOF) and tumor-bearing survival time after relapse (TST) in patients. RESULTS: The 1-, 2- and 3-year survival rate after operation in the WCA + CT group was 89.51%, 69.77% and 55.76% respectively, which was significantly higher than that in the CT group (83.86%, 59.33% and 49.43%) respectively (P < 0.05), but showed insignificant difference as compared with that in the WCA group (93.23%, 79.34% and 71.78%). Only 1-year metastasis in the WCA group was 15.25%, and in the WCA + CT group was 15.52%, the two were significantly lower than that in the CT group (35.48%, P < 0.05). But the comparison of 2-year metastasis rate among the 3 groups (28.81%, 41.38% and 45.16%) and 3-year metastasis rate among them (33.90%, 46.55% and 51.61%) were insignificantly different. The QOF and TST were markedly better in the WCA group than those in the CT group. CONCLUSION: WCA has preventive effect on relapse and metastasis in post-operational gastric cancer patients.