AIM: To study the antitumor effect of matrine in human hepatoma G2 (HepG2) cells and its molecular mechanism involved in antineoplastic activities. METHODS: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide...AIM: To study the antitumor effect of matrine in human hepatoma G2 (HepG2) cells and its molecular mechanism involved in antineoplastic activities. METHODS: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect viability of HepG2 cells. The effect of matrine on cell cycle was detected by flow cytometry. Annexin-V-FITC/PI double staining assay was used to detect cellular apoptosis. Cellular morphological changes were observed under an inverted phase contrast microscope. Transmission electron microscopy was performed to further examine ultrastructural structure of the cells treatedwith matrine. Monodansylcadaverine (MDC) staining was used to detect autophagy. Whether autophagy is blocked by 3-methyladenine (3-MA), an autophagy inhibitor, was evaluated. Expression levels of Bax and Beclin 1 in HepG2 cells were measured by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR).RESULTS: Matrine signif icantly inhibited the proliferation of HepG2 cells in a dose- and time-dependent manner, and induced G1-phase cell cycle arrest and apoptosis of HepG2 cells in a dose-dependent manner. The total apoptosis rate was 0.14% for HepG2 cells not treated with matrine. In contrast, the apoptosis rate was 28.91%, 34.36% and 38.80%, respectively, for HepG2 cells treated with matrine at the concentration of 0.5, 1.0 and 2.0 mg/mL. The remarkable morphological changes were observed under an inverted phase contrast microscope. Abundant cytoplasmic vacuoles with varying sizes were observed in HepG2 cells treated with matrine. Furthermore, vacuolization in cytoplasm progressively became larger and denser when the concentration of matrine was increased. Electron microscopy demonstrated formation of abundant autophagic vacuoles in HepG2 cells after matrine treatment. When the specif ic autophagic inhibitor, 3-MA, was applied, the number of autophagic vacuoles greatly decreased. MDC staining showed that the fluorescent density was higher and the number of MDC-labeled particles展开更多
The Qitianling granite batholith (QGB) is located in the southern Hunan Province, middle part of the Nanling Range, South China. Its total exposure area is about 520 km2. Based on our 25 single grain zircon U-Pb age d...The Qitianling granite batholith (QGB) is located in the southern Hunan Province, middle part of the Nanling Range, South China. Its total exposure area is about 520 km2. Based on our 25 single grain zircon U-Pb age data and 7 published data as well as the geological, petrological, and space distribution characteristics, we conclude that QGB is an Early Yanshanian (Jurassic) multi-staged composite pluton. Its formation process can be subdivided into three major stages. The first stage, emplaced at 163―160 Ma with a peak at about 161 Ma, is mainly composed of hornblende-biotite monzonitic granites and locally biotite granites, and distributed in the eastern, northern, and western peripheral parts of the pluton. The second stage, emplaced at 157―153 Ma with a peak at 157―156 Ma, is mainly composed of biotite granites and locally containing hornblende, and distributed in the middle and southeastern parts of the pluton. The third stage, emplaced at 150―146 Ma with a peak at about 149 Ma, is mainly composed of fine-grained (locally porphyritic) biotite granites, and distributed in the middle-southern part of the pluton. Each stage can be further disintegrated into several granite bodies. The first two intrusive stages comprise the major phase of QGB, and the third intrusive stage comprises the additional phase. Many second stage fine-grained granite bosses and dykes intruded into the first stage host granites with clear chilling margin-baking phenomena at their intrusive contacts. They were emplaced in the open fracture space of the earlier stage consolidated rocks. Their isotopic ages are mostly 2―6 Ma younger than their hosts. Conceivably, the time interval from magma emplacement, through cooling, crystallization, solidification, up to fracturing of the earlier stage granites cannot exceed 2―6 Ma. During the Middle-Late Jurassic in the Qitianling area and neighboring Nanling Range, the coeval granitic and basic-intermediate magmatic activities were widely developed. It indicates that the Early Yanshanian展开更多
基金Supported by National Natural Science Foundation of China, No. 30870364Science and Technology Support Program of Gansu Province, China, No. 0708NKCA129
文摘AIM: To study the antitumor effect of matrine in human hepatoma G2 (HepG2) cells and its molecular mechanism involved in antineoplastic activities. METHODS: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect viability of HepG2 cells. The effect of matrine on cell cycle was detected by flow cytometry. Annexin-V-FITC/PI double staining assay was used to detect cellular apoptosis. Cellular morphological changes were observed under an inverted phase contrast microscope. Transmission electron microscopy was performed to further examine ultrastructural structure of the cells treatedwith matrine. Monodansylcadaverine (MDC) staining was used to detect autophagy. Whether autophagy is blocked by 3-methyladenine (3-MA), an autophagy inhibitor, was evaluated. Expression levels of Bax and Beclin 1 in HepG2 cells were measured by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR).RESULTS: Matrine signif icantly inhibited the proliferation of HepG2 cells in a dose- and time-dependent manner, and induced G1-phase cell cycle arrest and apoptosis of HepG2 cells in a dose-dependent manner. The total apoptosis rate was 0.14% for HepG2 cells not treated with matrine. In contrast, the apoptosis rate was 28.91%, 34.36% and 38.80%, respectively, for HepG2 cells treated with matrine at the concentration of 0.5, 1.0 and 2.0 mg/mL. The remarkable morphological changes were observed under an inverted phase contrast microscope. Abundant cytoplasmic vacuoles with varying sizes were observed in HepG2 cells treated with matrine. Furthermore, vacuolization in cytoplasm progressively became larger and denser when the concentration of matrine was increased. Electron microscopy demonstrated formation of abundant autophagic vacuoles in HepG2 cells after matrine treatment. When the specif ic autophagic inhibitor, 3-MA, was applied, the number of autophagic vacuoles greatly decreased. MDC staining showed that the fluorescent density was higher and the number of MDC-labeled particles
基金Supported by National Natural Science Foundation of China (Grants Nos. 40730423, 40373014)Funds from the Ministry of Land and Resources (Grant No. 1212010632100)
文摘The Qitianling granite batholith (QGB) is located in the southern Hunan Province, middle part of the Nanling Range, South China. Its total exposure area is about 520 km2. Based on our 25 single grain zircon U-Pb age data and 7 published data as well as the geological, petrological, and space distribution characteristics, we conclude that QGB is an Early Yanshanian (Jurassic) multi-staged composite pluton. Its formation process can be subdivided into three major stages. The first stage, emplaced at 163―160 Ma with a peak at about 161 Ma, is mainly composed of hornblende-biotite monzonitic granites and locally biotite granites, and distributed in the eastern, northern, and western peripheral parts of the pluton. The second stage, emplaced at 157―153 Ma with a peak at 157―156 Ma, is mainly composed of biotite granites and locally containing hornblende, and distributed in the middle and southeastern parts of the pluton. The third stage, emplaced at 150―146 Ma with a peak at about 149 Ma, is mainly composed of fine-grained (locally porphyritic) biotite granites, and distributed in the middle-southern part of the pluton. Each stage can be further disintegrated into several granite bodies. The first two intrusive stages comprise the major phase of QGB, and the third intrusive stage comprises the additional phase. Many second stage fine-grained granite bosses and dykes intruded into the first stage host granites with clear chilling margin-baking phenomena at their intrusive contacts. They were emplaced in the open fracture space of the earlier stage consolidated rocks. Their isotopic ages are mostly 2―6 Ma younger than their hosts. Conceivably, the time interval from magma emplacement, through cooling, crystallization, solidification, up to fracturing of the earlier stage granites cannot exceed 2―6 Ma. During the Middle-Late Jurassic in the Qitianling area and neighboring Nanling Range, the coeval granitic and basic-intermediate magmatic activities were widely developed. It indicates that the Early Yanshanian
