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【Objective】 To investigate the role of RRM2 in prostate cancer and the mechanism. 【Methods】 The data of prostate cancer expression profile were downloaded from The Cancer Genome Atlas (TCGA). The correlation between RRM2 expression and clinicopathological features and prognosis of prostate cancer was analyzed. The protein expressions of RRM2 in 55 cases of prostate cancer and 38 benign tissues were determined with immunohistochemistry (IHC). The effects of RRM2 on the biological process of prostate cancer were assessed with bioinformatic analysis. The biological process of RRM2 affecting the progression of prostate cancer was verified with Western blot and flow cytometry. 【Results】 RRM2 was highly expressed in prostate cancer, and the expression was positively correlated with the clinical stage, pathological grade and metastasis of prostate cancer (P<0.05). Higher RRM2 expression predicted poorer survival. RRM2 co-expression positively correlated genes were involved in cell cycle pathways, pyrimidine nucleotide metabolism, and biological processes such as RNA transport. Cell cycle pathways were significantly enriched. RRM2 was highly correlated with CDK1 and PCNA molecules. RRM2 knockdown reduced the protein expressions of CDK1 and PCNA in DU145 and LNCap cell lines, which were arrested in the G2/M phase. 【Conclusion】 RRM2 promotes tumor progression by interfering with G2/M cycle of prostate cancer cells.
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OBJECTIVE Chronic kidney disease (CKD) has become a global public health problem with 10％-15％incidence rate, and inhibiting the renal interstitial fibrosis is considered to be a potential strategy to delay the progression of CKD. Z-Guggulsterone (Z-GS), an active compound from derived from Commiphora mukul, has been proved to be effective in various diseases. The present study aimes to determine the protective effect and the molecular mechanism of Z-GS on renal fibrosis. METHODS Unilateral ureteral obstruction (UUO) mice and hypoxia-induced HK-2 cells were used to simulate renal fibrosis in vitro and in vivo, respectively. The mice and cells were treated with different doses of Z-GS to observe the pharmacological action. Renal function, including Scr, BUN, and UA, were detected by commercial kits. H&E and Masson staining were performed to observe histopathological changes of kidney. Cell viability and LDH release of HK-2 cells were detected by commercial kits. Cell cycle distribution and apoptosis rate were analyzed by flow cytometry. Fibrosis markers were detected by immunohistochemistry and immunofluorescence analysis. Cell cycle related proteins and Klotho/p53 signaling were analyzed by Western blotting. RESULTS The results showed that Z-GS decreased the rise of Scr, BUN, and UA and lightened renal histopathological injury, which were induced by UUO. Besides, Z-GS administration alleviated renal fibrosis in mice by inhibiting the expressions of α-SMA, TGF-β and colla?genⅣ, and delayed G2/M cell cycle arrest by promoting the expressions of CDK1 and cyclinD1/B1 rate. Experiments in vitro indicated that Z-GS treatment significantly increased the cell viability while decreased the LDH release in hypoxia-induced HK-2 cells. In addition, hypoxia induced fibrosis and G2/M cycle arrest in HK-2 cells were retarded by Z-GS. The study of its possible mechanism exhibited that Z-GS treatment increased the level of Klotho and inhibited P53 level. Nev?ertheless, the effect of Z-GS on Klotho/P53 signaling was reversed by siRNA-Klotho. Moreover, siRNA-Klotho treatment eliminated the effects of Z-GS on G2/M cell cycle arrest and fibrosis. CONCLUSION This study clarified that Z-GS allevi?ated renal fibrosis and G2/M cycle arrest through Klotho/P53 signaling pathway. People who have suffered CKD may potentially benefit from treatment with Z-GS.
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Background: Several studies have shown that proton pump inhibitors (PPIs) can enhance the sensitivity of gastric cancer (GC) cells to chemotherapy and inhibit tumor proliferation and invasion. Aims: To investigate whether PPI could enhance chemosensitivity by inhibition of cell cycle-related genes in GC cells. Methods: Two human GC cell lines, AGS and HGC27 were treated with pantoprazole in different concentrations, and the cell viability was detected by CCK-8 assay. Transcriptome sequencing combined with KEGG enrichment analysis were used to determine the effect of PPI on cell cycle of GC cells, and the changes of cell cycle and its related genes were validated by flow cytometry, real-time PCR and Western blotting, respectively. Bioinformatics websites were employed to analyze the major differentially expressed cell cycle-related genes in GCs and their relationship with patients' prognosis. After transfection with FOXM1 plasmid or control plasmid, the inhibitory effect of PPI combined with cisplatin on GC cells was determined by CCK-8 assay. Results: PPI inhibited the proliferation of GC cells effectively in vitro. Transcriptome sequencing showed that the expression levels of G2/M phase-related genes, including FOXM1, PLK1, and AURKB were down-regulated in PPI-treated GC cells, and G2/M arrest was suggested by KEGG enrichment analysis. All these changes were proved by flow cytometry, real-time PCR and Western blotting. Bioinformatics analysis revealed that FOXM1, PLK1, and AURKB genes were highly expressed in GCs and correlated with a poor prognosis. The inhibitory effect of PPI combined with cisplatin on GC cells was superior to that of cisplatin alone, but could be partially reversed by overexpression of FOXM1. Conclusions: PPI treatment can induce G2/M arrest in GC cells by inhibiting cell cycle-related genes, and subsequently enhance the sensitivity of GC cells to chemotherapy.
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Glioblastoma multiforme (GBM) in the central nervous system is the most lethal advanced glioma and currently there is no effective treatment for it. Studies of sinomenine, an alkaloid from the Chinese medicinal plant,
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Objective: To investigate the effect of bisindolylmaleimide derivative L6 on inducing apoptosis of leukemia cells and its molecular mechanism. Methods: MTT assay was used to determinate the killing effect of L6 on HELL, K562, and KG1a cells. Flow cytometry was used to detect the effects of L6 on apoptosis, cell cycle, and differentiation of HEL cells. Western blotting was used to detect the expression of apoptosis-related protein. Finally, the effect of L6 on leukemia mouse was studied in vivo. Results: MTT assay showed that L6 had a stronger inhibitory activity against HEL, K562, and KG1a cell lines than the positive control PKC412 compound, with IC50 of (0.05 ± 0.03), (0.32 ± 0.01), and (0.19 ± 0.10) μmol/L, respectively. L6 could induce the apoptosis, G2/M arrest, megakaryocyte differentiation of HEL cells with a dose effect. Western blotting revealed that L6 mainly performed apoptosis by activating Caspase-3, which is an apoptotic executive protein. Hematoxylin-eosin (HE) staining of liver tissue of mice showed a reduction in HEL cell infiltration, but the more significant reduction in group L6 was observed, indicating that L6 could delay the metastasis of leukemia, and its effect was better than that of PKC412. Conclusion: Bisindolylmaleimide derivative L6 has a strong anti-leukemia activity, providing new hope for the development of new leukemia drugs.
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Objective: To study the effect of rocaglaol from Aglaia odorata on HepG2 proliferation and to explore the potential anti-tumor mechanism. Methods: The MTT, colony formation, EdU incorporation, and CFDA-SE assays were used to determine the anti-proliferative activity of rocaglaol in HepG2 cells. Apoptosis and cell cycle distribution effect induced by rocaglaol were carried out by flow cytometry. The effect of rocaglaol on protein involved in the G2/M checkpoint and the MAPK pathway were performed by Western blotting analysis. Results: Rocaglaol significantly inhibited the viability of HepG2 cells in a dose-dependent and time-dependent manner. Rocaglaol was more effective than doxorubicin in the growth inhibition of HepG2 cells. However, rocaglaol-induced cytotoxicity in normal human hepatic cell line L02 was lower than that of doxorubicin. Treatment with different concentrations of rocaglaol at 48 h caused G2/M cell cycle progression inhibition, rather than apoptosis in HepG2 cells. Rocaglaol can significantly reduce the expression of G2/M cell cycle-regulating proteins cdc25C, cdc2, and cyclin B1 as well as increase the expression of ERK and JNK phosphorylation levels. Further study found that U0126 can partly abrogate the anti-proliferative activity in HepG2 cells, G2/M phase arrest and the reduction in the protein expression levels of cdc2 and cdc25C induced by rocaglaol. Conclusion: Our results demonstrated that rocaglaol was superior to doxorubicin in the inhibition of HepG2 cells proliferation and the selectivity of L02 cell activity. We provided evidence that the rocaglaol had the ability to continuously over-activate the ERK signaling in HepG2 cells, leading to the inhibition of cell proliferation through G2/M phase arrest.
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Response gene to complement 32 (RGC-32) as an important response gene to complement was widely expressed in a lot of tissues and organs and participated in many biological processes such as cell proliferation and differentiation,cell cycle regulation,inflammation,immune regulation,and tumor,etc.As a cell cycle regulator,RGC-32 affected the development of numerous diseases by regulating the cell cycle.In recent years,many studies have shown that RGC-32 may be involved in the renal tubular injury and repair,and its role in the renal tubular injury and repair may be related to its regulation of cell cycle especially the G2/M.This article will make a brief review on the progress of the mechanism of RGC-32 regulating the renal injury and repair.
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Response gene to complement 32(RGC-32)as an important response gene to complement was widely expressed in a lot of tissues and organs and participated in many biological processes such as cell proliferation and differentiation, cell cycle regulation, inflammation, immune regulation, and tumor, etc.As a cell cycle regulator, RGC-32 affected the development of numerous diseases by regulating the cell cycle.In recent years, many studies have shown that RGC-32 may be involved in the renal tubular injury and repair, and its role in the renal tubular injury and repair may be related to its regulation of cell cycle especially the G2/M.This article will make a brief review on the progress of the mechanism of RGC-32 regulating the renal injury and repair.
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Objective: Marsdenia tenacissima extract (MTE) is a traditional Chinese herbal medicine with anti-cancer activity. In some previous studies, different mechanism actions of the anti-cancer effect of MTE have been revealed. In this study, we first observed that MTE exhibited G2/M cell cycle arrest on two different human breast cancer cell lines, MDA-MB-231 and MCF-7 by mediating 14-3-3σ and c-myc. Methods: The effect of MTE on G2/M cell cycle arrest was evaluated in MDA-MB-231 and MCF-7 cell lines. MTT assay was done for evaluation of cell viability. Flow cytometry was employed for cell cycle analysis. Western blotting analysis and immunohistochemistry were performed to analyze the expression of G2/M cell cycle-related key protein in cells and tissue samples. Animal studies have been conducted to elucidate the anti-tumor effect of MTE. Results: Cell cycle is the backbone for developing cancer. Cell cycle proteins play a major role in the progression of cell cycle and cell proliferation. However, some key protein directly or indirectly modulate the action of cell cycle protein that highly affect cell cycle regulation. In order to investigate cellular proliferation of cancer, we observed that MTE induced the upregulation of 14-3-3σ and downregulation of c-myc, and then reduced the expression of G2/M cell cycle associated key protein, leading to the inhibition of cellular entry into mitosis phase. We also confirmed that MTE exerted a significant antitumor effect on the MDA-MB-231 xenograft model in vivo. Conclusion: G2/M cell cycle arrest occurred by the action of MTE, mediated by the upregulation of 14-3-3σ as well as downregulation of c-myc in MDA-MB-231 and MCF-7 cell lines.
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According to the literatures at home and abroad, this paper comprehensively summarized and analyzed the research progress of regulation ways and means of various active ingredients of Chinese materia medica (CMM) on G2/M phase of tumor cells in recent years. Most of the active ingredients of CMM cause mitotic disaster of tumor cells by affecting Cyclin-CDK complex or inducing mitotic catastrophe in tumor cells, inducing DNA damage, mitotic defects and cytokinesis failure, thereby blocking tumor cells in G2/M phase, thus inhibiting tumor cell proliferation and finally inducing apoptosis. The active constituents of CMM can inhibit tumor growth and induce apoptosis by up-regulating or inhibiting key genes at the G2/M detection site, arresting tumor cells in G2/M phase, thereby exerting antitumor effects.
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OBJECTIVE: To investigate the apoptosis effect of human hepatocellular carcinoma cell line SMMC-7721 induced by dihydroartemisinin in vitro and the possible mechanism. METHODS: After treatment with 25, 50, 100, 200, and 400 μmol•L-1 dihydroartemisinin for 24 h. The proliferation inhibitory effect of dihydroartemisinin on SMMC-7721 cell was detected by MTT assay. Cell cycle and apoptosis were detected by flow cytometry. The change of apoptotic morphology was detected by confocal laser scanning microscopy. Rho 123 staining method was used to detect the changes of mitochondrial membrane potential. Western blot was used to detect expression of Bcl-2, Bax, Cleaved Caspase-3, Cleaved Caspase-9 and Cyto C. RESULTS: MTT results showed that 25-400 μmol•L -1 dihydroartemisinin can inhibit the proliferation of SMMC-7721 cells obviously. The cell cycle detection results of flow cytometry showed that dihydroartemisinin could block SMMC-7721 cell cycle in G2/M phase. The results of Hochest 333258 staining showed that the nuclei were heterogeneous, condensed and fragmented in the DHA treatment group. The cell apoptosis detection results of flow cytometry showed that the apoptosis rate of dihydroartemisinin treated groups were increased obviously (P < 0.01). The results of Rho 123 staining showed that the mitochondrial membrane potential was decreased significantly (P < 0.01). Western blot results showed that the expression of Bcl-2 was down-regulated, expression of Bax was up-regulated, the ration of Bax /Bcl-2 was increased and the expression of Cleaved Caspase-3, Cleaved Caspase-9 and Cyto C were up-regulated. CONCLUSION: Dihydroartemisinin can induce apoptosis of SMMC-7721 cells, on the mechanism of apoptosis may be related to mitochondrial pathway.
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AIM:To investigate the effect of fenbendazole (FBZ) on the proliferation of human chronic myelogenous leukemia (CML) cell line K562.METHODS:The CCK-8 assay was used to detect the effect of FBZ on viability of the K562 cells and normal peripheral blood mononuclear cells (PBMC).The cell growth was measured by the method of Trypan blue exclusion.The cell cycle was analyzed by flow cytometry.The cell cycle-related proteins were detected by Western blot.RESULTS:The growth of K562 was significantly inhibited by FBZ.However, it elicited little cytotoxic effect on PBMC.Furthermore, FBZ induced G2/M phase arrest and mitotic catastrophe in the K562 cells based on the changes of nuclear morphology, DNA content, mitotic marker analysis and the number of polykaryocytes.CONCLUSION:Fenbendazole significantly inhibits the proliferation of K562 cells and induces cell cycle arrest at G2/M phase by the regulation of cell cycle-related proteins.
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AIM:To investigate the mechanism of the radiosensitizing effect of maximum non-cytotoxic doses of tetrandrine (Tet) on nasopharyngeal carcinoma cell lines CNE1 and CNE2.METHODS:The cells were treated with maximum non-cytotoxic doses of Tet (for CNE1 cells at 1.5 μmol/L and for CNE2 cells at 1.8 μmol/L),irradiation at 4 Gy,or combination of irradiation and maximum non-cytotoxic doses of Tet.The cell cycle distribution was analyzed by flow cytometry.The protein levels of γ-H2AX,cleaved caspase-3,p-CDC25C,CDK1,p-CDK1,cyclin B1,ERK and p-ERK were determined by Western blot.RESULTS:The expression of γ-H2AX was increased in CNE1 cells and CNE2 cells after combined treatment with irradiation and maximum non-cytotoxic doses of Tet.The percentages of CNE1 cells and CNE2 cells at G2/M phase in irradiation group were (18.09 ±0.42)％ and (18.48 ± 1.32)％,respectively,which were decreased to (15.88 ± 1.04) ％ and (13.80 ± 0.82) ％ in combined treatment group,respectively (P ＜ 0.05).Combined treatment enhanced the increase in the protein level of cleaved caspase-3 caused by irradiation.The protein levels of pCDC25C and p-CDK1 were increased in a dose-dependent manner by Tet treatment (P ＜ 0.05),while the expression of CDK1 showed no difference among different doses of Tet treatments.The protein levels of p-CDC25C,p-CDK1 and CDK1 showed no difference after the treatment with maximum non-cytotoxic doses of Tet.The combined treatment with irradiation and the maximum non-cytotoxic doses of Tet decreased the protein levels of p-CDC25C and p-CDK1 (P ＜0.05),increased the expression of cyclin B1,and had no influence on the expression of CDK1 （ P ＜0.05).The combined treatment resulted in an increase in the protein level of p-ERK1 (P ＜ 0.05).CONCLUSION:The maximum non-cytotoxic doses of Tet enhance the DNA damage and apoptosis in CNE1 cells and CNE2 cells caused by irradiation,and the mechanism might be associated with ending of G2/M arrest via activation of ERK/CDC25C/CDK1/cyclin B1 pathways.
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AIM:To investigate the mechanism of the radiosensitizing effect of maximum non-cytotoxic doses of tetrandrine (Tet) on nasopharyngeal carcinoma cell lines CNE1 and CNE2.METHODS:The cells were treated with maximum non-cytotoxic doses of Tet (for CNE1 cells at 1.5 μmol/L and for CNE2 cells at 1.8 μmol/L),irradiation at 4 Gy,or combination of irradiation and maximum non-cytotoxic doses of Tet.The cell cycle distribution was analyzed by flow cytometry.The protein levels of γ-H2AX,cleaved caspase-3,p-CDC25C,CDK1,p-CDK1,cyclin B1,ERK and p-ERK were determined by Western blot.RESULTS:The expression of γ-H2AX was increased in CNE1 cells and CNE2 cells after combined treatment with irradiation and maximum non-cytotoxic doses of Tet.The percentages of CNE1 cells and CNE2 cells at G2/M phase in irradiation group were (18.09 ±0.42)％ and (18.48 ± 1.32)％,respectively,which were decreased to (15.88 ± 1.04) ％ and (13.80 ± 0.82) ％ in combined treatment group,respectively (P ＜ 0.05).Combined treatment enhanced the increase in the protein level of cleaved caspase-3 caused by irradiation.The protein levels of pCDC25C and p-CDK1 were increased in a dose-dependent manner by Tet treatment (P ＜ 0.05),while the expression of CDK1 showed no difference among different doses of Tet treatments.The protein levels of p-CDC25C,p-CDK1 and CDK1 showed no difference after the treatment with maximum non-cytotoxic doses of Tet.The combined treatment with irradiation and the maximum non-cytotoxic doses of Tet decreased the protein levels of p-CDC25C and p-CDK1 (P ＜0.05),increased the expression of cyclin B1,and had no influence on the expression of CDK1 （ P ＜0.05).The combined treatment resulted in an increase in the protein level of p-ERK1 (P ＜ 0.05).CONCLUSION:The maximum non-cytotoxic doses of Tet enhance the DNA damage and apoptosis in CNE1 cells and CNE2 cells caused by irradiation,and the mechanism might be associated with ending of G2/M arrest via activation of ERK/CDC25C/CDK1/cyclin B1 pathways.
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<p><b>OBJECTIVE</b>To explore the role of p21 in ionizing radiation-induced changes in protein levels during the G2/M transition and long-term G2 arrest.</p><p><b>METHODS</b>Protein expression levels were assessed by western blot in the human uveal melanoma 92-1 cells after treatment with ionizing radiation. Depletion of p21 was carried out by employing the siRNA technique. Cell cycle distribution was determined by flow cytometry combined with histone H3 phosphorylation at Ser28, an M-phase marker. Senescence was assessed by senescence- associated-β-galactosidase (SA-β-gal) staining combined with Ki67 staining, a cell proliferation marker.</p><p><b>RESULTS</b>Accompanying increased p21, the protein levels of G2/M transition genes declined significantly in 92-1 cells irradiated with 5 Gy of X-rays. Furthermore, these irradiated cells were blocked at the G2 phase followed by cellular senescence. Depletion of p21 rescued radiation-induced G2 arrest as demonstrated by the upregulation of G2/M transition kinases, as well as the high expression of histone H3 phosphorylated at Ser28. Knockdown of p21 resulted in entry into mitosis of irradiated 92-1 cells. However, cells with serious DNA damage failed to undergo cytokinesis, leading to the accumulation of multinucleated cells.</p><p><b>CONCLUSION</b>Our results indicated that p21 was responsible for the downregulation of G2/M transition regulatory proteins and the bypass of mitosis induced by irradiation. Downregulation of p21 by siRNA resulted in G2-arrested cells entering into mitosis with serious DNA damage. This is the first report on elucidating the role of p21 in the bypass of mitosis.</p>
Subject(s)
Humans , Cell Cycle Checkpoints , Radiation Effects , Cell Line, Tumor , Cyclin-Dependent Kinase Inhibitor p21 , Genetics , Metabolism , DNA Damage , Down-Regulation , Fibroblasts , Metabolism , Radiation Effects , Gene Expression Regulation , Radiation Effects , Mitosis , Radiation Effects , RNA Interference , RNA, Small Interfering , Radiation, Ionizing , Up-RegulationABSTRACT
OBJECTIVE@#To identify the influence on radiosensitivity of lung glandular cancer cells when excisions repair cross-complementing group1 (ERCC1) gene was silenced by targeted siRNA.@*METHODS@#siRNA which targeting to ERCC1 and control siRNA was designed and synthesized. The human lung glandular cancer SPC-A-1 cells was transfected. A total of 56 nude mice were divided into two groups, and two kinds of SPC-A-1 cells were transplanted to armpit of right forelimb, to establish the nude mice subcutaneous xenotransplanted tumor model of human lung glandular cancer cells. After the tumor was developed, the nude mice were randomly divided into four groups and accepted different doses of X-Ray radiation, then the change of tumor volume, survival time of mice in every group were recorded and the average lifetime was calculated. Twenty-one days later of X-ray experiment, two mice were taken and killed in each group and the tumors organizations were stripped. The cell apoptosis rate and cell cycle distributions were obtained by FCM (flow cytometry).@*RESULTS@#The volume of tumor which ERCC1 gene was silenced was less than single irradiation group after X-ray irradiation, and the growth speed was slower and the lifetime of mice was lengthened as well (P < 0.05). The cells apoptosis rate and the rate of G2/M cells which ERCC1 gene was silenced were higher than the same dose control group and the rate of G1 cells were lower, which indicated that the cells could be stopped at G2/M point, the cell proliferation was inhibited, the cell apoptosis was promoted and the radiation sensitivity was improved after the ERCC1 was silenced.@*CONCLUSIONS@#The radiation sensitivity of lung glandular tumor could be improved after the ERCC1 gene was silenced by siRNA.
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Aim To investigate the effects of diallyl di-sulfide( DADS) on G2/M arrest in Chk1/MGC803 and Chk2/MGC803 cells so as to establish stable human gastric cancer MGC803 cells with overexpression of Chk1/2 gene. Methods The colony formation, flow cytometry, RT-PCR and Western blot were used to de-tect the proliferation, cell cycle, and expression of Chk1/2 mRNA and protein, p-Chk1/2, CDC25C and cyclinB1, respectively. Results The colony formation showed that the colony forming efficiency in Chk1/MGC803 and Chk2/MGC803 cells treated by 30 mg· L-1 DADS was lower than in control group and vector group ( P <0. 05 ) . Flow cytometry demonstrated that 41. 3%, 57. 4%, 68. 9% and 42. 9% of G2/M cells in Chk1/MGC803 were increased than in MGC803 and Chk2/MGC803 , respectively after treated by DADS in 12,24, 36 and 48 h(P <0. 05). At the same time, RT-PCR disclosed that expression of Chk1 and Chk2 mRNA had no marked change. Western blot showed that total proteins of Chk1 and Chk2 and p-Chk2 had invisible change, but expression of p-Chk1 was up-reg-ulated, and CDC25C and cyclinB1 were down-regula-ted time-dependently in Chk1/MGC803 cells ( P <0. 05 ) . Conclusion DADS arrests MGC803 cells at G2/M by increasing p-Chk1 expression to cause down-regulation of CDC25C and cyclinB1 simultaneously.
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Aim To investigate the anti-tumor effects of FS-108 an Hsp90 inhibitor, on oncogene addicted EBC-1 and A375 cells. Methods SRB assay was performed to investigate cell proliferation. Immunoblot was conducted to investigate the specific proteins. FACS was conducted to test cell cycle distribution and apoptosis. Transwell assay was conducted to investigate cell motility. Results FS-108 significantly suppressed cell proliferation of EBC-1 and A375 cancer cells with IC50 at 25. 53 nmol · L-1 and 30. 02 nmol · L-1 re-spectively. FS-108 treatment triggered the degradation of key client proteins such as c-Met and B-Raf and thereby reduced their downstream AKT and ERK signa-ling pathways. The FACS analysis results demonstrated that FS-108 treatment induced G2/M phase arrest and apoptosis significantly. Furthermore, FS-108 inhibited the migration of EBC-1 and A375 cells. Conclusion As a potent Hsp90 inhibitor, FS-108 can inhibit onco-gene addicted cancer cells proliferation through induc-tion of G2/M phase arrest and apoptosis.
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To investigate the anti-hepatoma mechanism of α-pinene, HepG2 cell was treated with α-pinene and the change of cell cycle was examined by flow cytometry. The expression of miR-221, which was related the regulation of G₂/M phase, was detected by quantitative Real-time PCR. Meanwhile, TargetScan and other online bioinformatics methods were used to analyze and predict the target genes of miR-221, then the expression level of related target genes were detected by quantitative Real-time PCR. The results showed that α-pinene inhibited the proliferation of HepG2 cells in dose-dependent manner. It was also proved that HepG2 cells were arrested at G₂/M phase by α-pinene (P<0.05). The expression of miR-221 was down-regulated in α-pinene treated HepG2 cell. The bioinformatics analysis showed that CDKN1B/P27 and CDKN1C/P57 may be the protential targets of miR-221 and both of them were significantly up-regulated（P<0.001,P<0.05）by α-pinene treatment. According to these results, it was believed that α-pinene may inhibit the proliferation of hepatocellular carcinoma cells through arrest the cell at G₂/M phase, which may be associated with the down-regulate of the miR-221 expression and up-regulate of the CDKN1B/P27 and CDKN1C/P57 expression.
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The aim of the study was to investigate the anti-proliferation and apoptosis-inducing effects of S1, a novel tetrandrine derivative, in human gastric cancer BGC-823 cells and explore the possible mechanism of action. The anti-proliferative activity was determined by MTT assay; the induction of cell cycle arrest and apoptosis were detected by flow cytometry. Quantitative real time RT-PCR and Western blotting were used to evaluate the mRNA and protein expression levels in mitochondrial pathway. S1 significantly reduced cell viability and induced a G2/M phase arrest and apoptosis in dose- and time-dependent manner. Further studies showed that S1 increased mRNA and protein expression of Bax and the Bax/Bcl-2 ratio. Moreover, S1 decreased the protein expression of procaspase-9 and procaspase-3, suggesting that the induction of apoptosis may be related to the alteration of the ratio of Bax/Bcl-2 and the activation of caspases. These findings suggested that S1 merits further investigation as a novel therapeutic agent for the treatment of human gastric cancer.