Cancer Cell Lines SMMC-7721 Research Articles

Investigation on the substitution effects of the flavonoids as potent anticancer agents: a structure–activity relationships study

Three series of flavonoid analogues substituted with different aminomethyl substitutions at C-6, C-7, and C-8 were designed and synthesized for the structure–activity relationship studies as potent anticancer agents. The prepared analogues were evaluated for their in vitro inhibitory activity against the growth of the hepatic cancer cell lines HepG2 and SMMC-7721. Structure–activity relationships indicated that not only the compounds with amino methyl groups were more active than those without the groups in the same series but also the compounds substituted by aminomethyl groups at position C-8 were more active than those at positions C-6 and C-7.

Design, synthesis and biological evaluation of novel 4-thiazolidinones containing indolin-2-one moiety as potential antitumor agent

A series of novel 4-thiazolidinone and indolin-2-one hybrid derivatives 5a– 5s and 10a– 10s have been designed and synthesized and their cytotoxic activities were evaluated in vitro against three human cancer cell lines including HT-29 (human colon cancer), H460 (human lung cancer), MDA-MB-231 (human breast cancer) by MTT assay. Several potent target compounds ( 5m, 5p, 5s, 10a, 10c– 10g, 10m, 10p) were further evaluated against one cancer cell line SMMC-7721 (human liver cancer) and one normal cell line WI-38 (human fetal lung fibroblasts). Most of the prepared compounds exhibited significant antitumor activities against different human cancer cell lines. Compound 10c (IC 50 = 0.025 μM, 0.075 μM, 0.77 μM, 1.95 μM) was 52, 36, 4.8 and 3.3 times more active than Sunitinib (IC 50 = 1.3 μM, 2.7 μM, 3.7 μM, 6.47 μM) against HT-29, H460, MDA-MB-231 and SMMC-7721 cancer cell line, respectively.

A New Tetracyclic Diterpene from Jatropha curcas

AbstractJatrophodione A (1), a new diterpene with four rings, together with nine known compounds, caniojane (2), jatropholone A (3), jatropholone B (4), jatrogrossidione (5), 2‐epijatrogrossidione (6), heudelotinone (7), gossweilone (8), (3α)‐3‐hydroxy‐ent‐pimara‐8(14),15‐dien‐12‐one (9), and 12‐hydroxy‐13‐methylpodocarpa‐8,11,13‐trien‐3‐one (10), was isolated from the aerial parts of Jatropha curcas. Compounds 5, 6, 9, and 10 were found for the first time in this plant. Their structures were established by spectroscopic analysis, including 2D‐NMR spectroscopic techniques. Cytotoxicities of compounds 1, 2, 7, 8, and 9 were tested on the three cancer cell lines A549, Hela, and SMMC‐7721. Results showed that 7 exhibited cytotoxicity against SMMC‐7721 with an IC50 value of 21.68 μM, whereas 7 and 8 were active against A549 with the IC50 values of 16.04 and 20.47 μM, and against Hela with the IC50 values of 10.67 and 22.83 μM, respectively.

Study on antitumor activity of metal-based diketone complexes

The biological activity of diketone complexes—M(acac) (Acac = MeCOCHCOMe, M = Mo6+, and Co2+) in potential antitumor application was evaluated. The cancer cell lines SMMC-7721 (liver cancer) and SK-OV-3 (ovary cancer) were tested for their viabilities by the MTT method in vitro, which showed that the two compounds exhibited a significant activity against two cell lines. Co(acac)2 exhibited higher antitumor activity than MoO2(acac)2 did, which is attributed to the fact that Co(acac)2 could interact with λ-DNA and MoO2(acac)2 could not. The results showed that Co(acac)2 could be intercalated into the double helix of λ-DNA.

The Cyclooxygenase-2 Inhibitor Celecoxib Attenuates Hepatocellular Carcinoma Growth and c-Met Expression in an Orthotopic Mouse Model

To demonstrate in vivo tumor growth inhibition, the liver cancer cell lines HepG2, BEL7402, and SMMC7721 were independently inoculated into the livers of 45 6-week-old nude mice. After 24 h, mice were randomly divided into celecoxib (intragastric celecoxib suspension, 300 mg/kg), negative control (equal volume intragastric saline), and positive control (intraperitoneal injection of 6 mg/kg doxorubicin) and treated once per day for 3 days. Body weights, tumor diameters, and tumor expressions of proliferating cell nuclear antigen (PCNA) and c-Met were determined at 23 days posttreatments. Significant increases in body weight were observed in celecoxib- or doxorubicin-treated mice compared to saline-treated animals and tumor growth was significantly attenuated, accompanied by downregulation of tumor PCNA expression (p < 0.01). Weight gain, attenuated tumor growth, and reduced PCNA expression were similar following celecoxib or doxorubicin treatment. Celecoxib also significantly reduced c-Met expression in HepG2- and BEL7402-induced tumors, but not SMMC7721-induced tumors (p < 0.05). In conclusion, celecoxib effectively suppressed the in vivo growth of liver cancer in an orthotopic tumor model. Celecoxib also inhibited tumor cell PCNA expression independent of changes in c-Met expression, with some variability between different implanted cell lines. This preclinical demonstration of celecoxib efficacy and safety provides a foundation for future clinical investigations involving use of this agent alone or as a component of chemotherapeutic regimens for treatment of HCC.

Neoclerodane Diterpenoids fromScutellaria barbata

Eight new neoclerodane diterpenoids (1- 8), scutebatas H-O, together with eight known compounds, have been isolated from the whole plant of Scutellaria barbata D. Don. Their structures were elucidated on the basis of spectroscopic studies. In vitro cytotoxicity of selected compounds against cancer cell lines HL-60, SMMC-7721, A-549, MCF-7, and SW480 was evaluated. Compounds 5, 6, 7 showed moderate cytotoxicities against several human cancer cell lines, with IC50 values ranging from 12.6-31.4 µM. In addition, compound 1 showed selective cytotoxicity against MCF-7.

New Potential Anticancer Agent of Carborane Derivatives: Selective Cellular Interaction and Activity of Ferrocene-Substituted Dithio-o-carborane Conjugates

The large diversity of structures and unique bonding modes of organometallic complexes make them possible to act as promising candidate therapeutic agents. In this study, the new type of ferrocene-substituted dithio-o-carborane conjugates (FcSB1, FcSB2, and FcSBCO) has been synthesized, and their in vitro antineoplastic activities have been explored by means of the electrochemical study, the real time cell electronic sensing (RT-CES) system, and biological assays. The conjugate-cell interactions were first monitored by electrochemistry, and the results show different cell uptake efficiency for FcSB1, FcSB2, and FcSBCO toward target cells. Both the highly hydrophobic ferrocenyl and carboranyl groups render the conjugates able to rapidly enter cells and exert acute cytotoxicity after 4 h incubation in serum-free media. However, FcSB1, FcSB2, and FcSBCO display different inhibition efficiencies toward SMMC-7721 and HepG2 cancer cells via the G(0)/G(1) arrest mechanism in a physiological environment. The anticancer activity is in the order FcSB2 > FcSB1 > FcSBCO, which is parallel to the order of the redox potentials of the ferrocenyl groups in the three complexes. In particular, FcSB1 and FcSB2 display a potent selective inhibition effect on the proliferation of the cancer cell lines SMMC-7721 and HepG2, but almost no effect on the normal cell line, the human embryonic lung fibroblast (HELF) cells. Thus, these results may provide some clues for use of the ferrocene-carborane conjugates in developing anticancer drugs.

Synthesis and biological evaluation of cyclopenten-1-one Mannich base oxovanadium compound

2-Hydroxy-3-methyl-2-cyclopenten-1-one Mannich base oxovanadium compound (CPD-VO) was synthesized. The human cancer cell lines SMMC-7721 (liver cancer) and SK-OV-3 (ovary cancer) were tested for their viability by MTT in vitro, which showed that CPD-VO exhibited a remarkable activity against the two cell lines, with IC50 values <0.1 8 and <0.23 μg/ml, respectively. And clonogenic assays showed that CPD-VO at 6.250 μg/ml inhibited colony formation of SMMC-7721 cells by 99.10%. Also, CPD-VO suppressed tumor growth of Hep-A-22 (mouse liver cancer) in mice in vivo. Moreover, the interaction of CPD-VO with λ-DNA was investigated using UV–vis spectroscopy and viscosity. The results showed that CPD-VO was intercalated into the double-helix of λ-DNA. In flow cytometry analysis, the ratio of apoptotic cells was up to 8.15% after treatment with CPD-VO at 2.34 μg/ml after 30 min, suggesting that the antitumoral activity of CPD-VO came from activation of the apoptotic pathway. CPD-VO is a promising antitumoral agent.

Damages within coding region of mitochondrial DNA in human liver cancer cell line SMMC-7721 exposured to X-ray

Objective:To study the damages within the coding region of the mitochondrial DNA in a human liver cancer cell line SMMC-7721 after exposure to different doses of X-ray irradiation.Methods:SMMC-7721 cells were radiated by different do- ses of X-ray(0,20,30,and 50 Gy).Primers for multiple PCR detection were designed according to the sequences of the coding region of human mitochondrial DNA,and the degree and location of mitochondrial DNA breaks were examined by LM-PCR (ligation mediated-PCR)and gene scanning technique.Results:The results of gene scanning showed the existence of X-ray sen- sitive breakpoints in the mitochondrial DNA coding region in human liver cancer cell line SMMC-7721 ,the degree of damage in- creased with the irradiation doses;the sensitive breakpoints were not distributed at random.The damage of heavy strand was se- vere than that in the light strand.Conclusion:X-ray irradiation can cause obvious damage to the coding region of the mitochon- drial DNA in human liver cancer cell line SMMC-7721,and the changes of mitochondrial DNA may play a role in the clinical therapy of cancer.

Combined anti-tumor effects of mDRA-6 and nimesulide on human hepatocellular cancer cell line SMMC-7721

The aim of this work was to evaluate anti-tumor effects of mDRA-6 plus nimesulide on a human hepatocellular cancer cell line, SMMC-7721, and study the main mechanisms. The DR5 receptor of SMMC-7721 cells was detected by flow cytometry (FCM). For further experimental application, SMMC-7721 cells were treated with proper dose of mDRA-6, nimesulide, or mDRA-6 plus 200 μmol/L nimesulide; untreated SMMC-7721 cells were comparably set as control. Cytotoxicity was tested by MTT assay; cell morphology was examined using Hoechst 33258 staining; and apoptosis was determined by FCM. The positive rate of DR5 on SMMC-7721 was 95.0%. Either mDRA-6 or nimesulide alone induces SMMC-7721 cell death in a dose-dependent manner. Treatment of 1,600 ng/mL mDRA-6 for 12h led to a cell-death rate of 35.0%, while an increased cell-death rate (91.1%) was found under the same condition of mDRA-6 treatment supplemented with 200 μmol/L nimesulide. Hoechst 33258 and Annexin V/PI staining confirmed apoptosis as the main cause of this anti-tumor response. Both mDRA-6 and nimesulide can induce apoptosis of SMMC-7721 cells, and they have synergistic anti-tumor activities against SMMC-7721.

Mechanism for decrease of glucocorticoid receptor mRNA in human hepatic cancer cell line SMMC-7721 during heat shock

Objective:To investigate the mechanism for decrease of glucocorticoid receptor mRNA(GR mRNA)during heat shock response.Methods:The changes of GR mRNA level in SMMC-7721 cells were examined by semi-quantitative RT-PCR during heat shock response.We designed 2 RNA fragments paired with GR gene intron E and cross exon 3,4,and 5,which were used as probes for in situ hybridization with the sequences in SMMC-7721 cells at different periods after heat shock.The result was subjected to confocol microscope observation and computer image analysis.The transcription,degradation,and splicing of GR mRNA were investigated.Results:RT-PCR showed that the GR mRNA level was decreased during heat shock response.In situ hybridization revealed that both GR mRNA and GR pre-mRNA levels were lower in the heat shock group than in the non-heat shock group(P0.05).GR pre-mRNA level was higher in cells treated with actinomycine D before heat shock than in cells only treated with actinomycine D,while the GR mRNA level was lower in cells treated with actinomycine D before heat shock(P0.05).Conclusion:During heat shock response,GR mRNA transcription is suppressed,the splicing of GR pre-mRNA is suppressed,and GR mRNA degradation is accelerated.

Synthesis and biological evaluation of decavanadate Na 4Co(H 2O) 6V 10O 28·18H 2O

We have reported the synthesis and biological evaluation of a decavanadate Na 4Co(H 2O) 6V 10O 28·18H 2O (CoV 10) designed as a potential antitumoral agent. The human cancer cell lines SMMC-7721 (liver cancer) and SK-OV-3 (ovary cancer) were tested for their viability by the MTT method in vitro, which showed that the compound exhibited a remarkable activity against two cell lines with IC 50 values smaller than 0.24 μg/mL, 0.32 μg/mL, respectively. CoV 10 showed the tumor growth suppression for Hep-A-22 (mice liver cancer) in tumor bearing mice in vivo. In addition, using flow cytometry analysis, the ratio of apoptotic cells was up to 8.33% with treatment of CoV 10 at 1.56 μg/mL after 30 min, suggesting that the antitumoral activity of CoV 10 comes from the activation of the apoptotic pathway.

23,24-Dihydrocucurbitacin B induces G 2/M cell-cycle arrest and mitochondria-dependent apoptosis in human breast cancer cells (Bcap37)

23,24-Dihydrocucurbitacin B (DHCB), a cucurbitacin-derived compound known to posses anticancer and anti-inflammatory activities. In this study, DHCB, isolated from roots of Trichosanthes kirilowli which is a traditional Chinese herb medicine used as treatments for cancer and other diseases, has been found to inhibit the proliferation of human cancer cell lines Bcap37, HeLa, SW620, SMMC-7721, K562 and MCF-7 in a dose- and time-dependent manner, and induce apoptosis in human breast cancer cell line Bcap37 at low concentration. DHCB-induced Bcap37 apoptosis was characterized with the changes in nuclear morphology, DNA fragmentation, activation of caspase-like activities, poly(ADP-ribose) polymerase cleavage, release of cytochrome c into cytosol. The cell death was partly prevented by a caspase-family inhibitor Z-VAD-FMK. The results suggest that DHCB-induced Bcap37 apoptosis through mitochondrial dependent pathway. Flow cytometric analysis revealed that at the lower dose of 1.8 and 3.6 μM, DHCB-induced cancer cell lines death via an apoptotic process rather than necrotic one; whereas, the higher dose of 8.9, 17.9 and 35.7 μM induced cell death via the necrotic process. Cell-cycle analysis demonstrated DHCB induction of G 2/M phase cell-cycle arrest and apoptosis. The overall results suggest that DHCB might have the therapeutic value against human cancer cell lines, especially the breast cancer cell lines.

Steep pulsed electric fields modulate cell apoptosis through the change of intracellular calcium concentration

A steep electric pulsed field with low intensity (150–250 V/cm) and relative long time (10 min) was applied to adherent liver cancer cell line SMMC-7721 and the liver cell line HL-7702. Results showed that the electric field with intensity of 200 and 250 V/cm could trigger cell apoptosis, whereas the SMMC-7721 cell was more sensitive to the electric stimulation than the HL-7702 cell. Laser Scanning Confocal Microscope (LSCM) was used to measuring the real-time change of cytosolic free Ca 2+ concentration. When cells were exposed electric pulses with 100 V/cm intensity for 10 min, there was no significant change of intracellular calcium concentration. With the intensity increased to 200 and 250 V/cm, intracellular calcium concentration decreased significantly. Results demonstrated the relationship between the apoptosis and change of intracellular calcium concentration. And the steep electric pulsed field can be used to the cancer therapy.

Construction of recombinant eukaryotic expression vector for human endostatin and its expression in liver cancer cell line SMMC-7721

Construction of recombinant eukaryotic expression vector for human endostatin and its expression in liver cancer cell line SMMC-7721

Antisense Tcf inhibits the neoplastic growth of liver cancer cells.

T cell transcription factors are nuclear effectors of the Wnt signaling transduction pathway and play crucial roles in embryonic and malignant development. Our previous study showed increased expression level of Tcf mRNA in liver cancer. In the present paper, antisense Tcf RNA was used to explore the possible therapeutic effect on liver cancer cells by interrupting the abnormal Wnt pathway. Antisense expression vectors containing the conserved sequence of Tcf cDNA were constructed and transfected into a human liver cancer cell line SMMC-7721. Tumorigenic potential was determined by cellular growth assay and tumor growth in nude mice. The stable transfection of anti-sense Tcf in SMMC-7721 cells significantly reduced Tcf expression at both mRNA and protein levels compared with parental and mock-transfected cells. Antisense-mediated suppression of Tcf inhibited the in vitro proliferation and in vivo tumor formation ability. Furthermore, the apoptosis rate of antisense transfected cells was significantly higher than that of control, indicating that antisense RNA suppressed malignant growth by induction of apoptosis. Our studies demonstrate the critical role of Wnt signaling pathway in the neoplastic growth of liver cancer cells and suggest that inhibition of Tcf activity with antisense Tcf RNA may be a potential new gene therapy method for liver cancer.

Potential involvement of leptin in carcinogenesis of hepatocellular carcinoma.

To investigate the potential involvement of leptin in carcinogenesis of hepatocellular carcinoma (HCC) and to elucidate the etiology, carcinogenesis and progress of HCC. Expressions of Ob gene product, leptin and its receptor, Ob-R were investigated in 36 cases of HCC specimens and corresponding adjacent non-tumorous liver tissues with immunohistochemical staining. The effect of leptin on proliferation of Chang liver cell line and liver cancer cell line SMMC-7721 was studied with cell proliferation assay (MTT). Leptin expression was detected in 36 cases of adjacent non-tumorous liver tissues (36/36, 100%) with moderate (++) to strong (+++) intensity; and in 72.22%(26/36) of HCC with weaker (+) intensity (P<0.05). Thirty of 36 (83.33%) cases of adjacent non-tumorous liver tissues were positive for Ob-R, with moderate (++) to strong (+++) intensity. In HCC, 11/36 (30.56%) cases were positive, with weak (+) intensity (P<0.05). In cell proliferation assay, leptin inhibited the proliferation of Chang liver cells. The cell survival rate was 10-13% lower than that of the untreated cells (P>0.05). Leptin had little effect on the proliferation of liver cancer cells (P>0.05). High level expression and decreased or absent expression of leptin and its receptor in adjacent non-tumorous liver cells and HCC cells, inhibitory effect of leptin on the proliferation of normal Chang liver cells and no effect of leptin on proliferation of liver cancer cells, may provide new insights into the carcinogenesis and progression of human HCC. It could be assumed that leptin acting as an inhibitor and/or promoter, is involved in the process of carcinogenesis and progress of human HCC.

Construction and biological effects in vitro of double-directed tissue-specific HSV-tk/GCV anti-tumor system

Objective: To construct a specifically targeted gene delivery and expression system, and to investigate the special killing effect of the HSV-tk/GCV system on human liver cancer cellsin vitro.Methods: The anti-transferrin receptor (TfR) ScFv-GAL4 fusion protein expression vector ScFv-GAL4-pET28a and the eukaryotic expression plasmid pEBAF/tk-GAL4rec were constructed by recombinant DNA technology. After the induction by IPTG, we obtained the anti-TfR ScFv-GAL4 fusion protein as delivery vector to transfect pEBAF/tk-GAL4rec into the human liver cancer cell lines HepG2 and SMMC7721 and the human lung cancer cell line A549 that overexpress TfR via receptor-mediated endocytosis. The positive cell clones were selected by hygromycin and named HepG2/tk, SMMC7721/tk and A549/tk, respectively. Cell killing after GCV application was determined by MTT.Results: The correct structure of the ScFv-Gal4 fusion protein and the plasmid pEBAF/tk-GAL4rec was confirmed by double enzyme digestion, SDS-PAGE and sequencing. HepG2/tk cells that express alphafetoprotein (AFP) to high levels (845 ng/ml) were very sensitive to GCV, while SMMC7721/tk cells that express AFP at low levels (2 ng/ml) and AFP-negative A549/tk cells were only slightly or not sensitive to GCV.Conclusion: The double-directed and tissue-specific HSV-tk/GCV anti-tumor system shows good targeting to tumor cells.

Apoptosis of human liver cancer cell line SMMC-7721 induced by Senjiu capsule

Apoptosis of human liver cancer cell line SMMC-7721 induced by Senjiu capsule