肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2014年
9期
587-591
,共5页
肝肿瘤%白细胞介素24%核心蛋白聚糖%细胞,HepG2%基因疗法%细胞增殖%凋亡%细胞周期阻滞
肝腫瘤%白細胞介素24%覈心蛋白聚糖%細胞,HepG2%基因療法%細胞增殖%凋亡%細胞週期阻滯
간종류%백세포개소24%핵심단백취당%세포,HepG2%기인요법%세포증식%조망%세포주기조체
Liver neoplasms%Interleukin-24%Decorin%Cell,HepG2%Gene therapy%Cell proliferation%Apoptosis%Cell cycle arrest
目的 观察外源基因重组人源白细胞介素(IL)-24(rhIL-24)联合重组人源核心蛋白聚糖(rhDCN)对人类肝细胞癌HepG2细胞的增殖、凋亡和细胞周期的影响.方法 采用脂质体瞬时转染法将pcDNA3.1(+)-IL-24和pcDNA3.1(+)-DCN质粒转染至HepG2细胞,48 h后倒置显微镜下观察细胞生长状况、形态学改变.分别在转染后24h、48 h和72 h,采用四甲基偶氮唑蓝(MTT)法观察IL-24和DCN单独及联合转染对HepG2细胞的增殖抑制效应.转染后48 h流式细胞术检测HepG2细胞凋亡情况,并进行细胞周期阻滞分析.结果 与对照组相比,转染目的基因48 h后显微镜下可见细胞生长不同程度地被抑制,并可见典型的凋亡细胞形态改变,以联合转染组改变更为明显.MTT结果显示转染后48 h和72 h,双基因联合转染组的抑制率分别是(31.88±6.57)％和(36.83±3.76)％,与对照组和单独转染组比较,差异均有统计学意义(P＜0.01).流式细胞术结果显示,单独转染IL-24和DCN基因可诱导HepG2细胞出现不同程度凋亡,而双基因联合转染组细胞凋亡率可达(32.56±0.90)％,与空质粒转染组的(2.98±0.72)％、空白细胞对照组的(3.50±0.92)％、IL-24组的(20.01±1.08)％和DCN组的(22.20±0.91)％比较,差异均有统计学意义(P＜0.01).细胞周期分析结果表明,与对照组相比,IL-24基因转染组的G2/M期和DCN基因转染组的G0/G1期细胞比例明显增高分别为(11.24±0.35)％、(77.93±0.67)％,而双基因联合转染组G0/G1期和G2/M期细胞比例同时增高,分别是(71.36±0.60)％和(10.39±0.67)％,差异有统计学意义(P＜0.01).结论 联合转染IL-24和DCN基因可以协同发挥较单独应用更强的抑制HepG2细胞增殖和诱导HepG2细胞凋亡的作用.IL-24可使HepG2细胞发生G2/M期阻滞,DCN可使HepG2细胞发生G0/G1期阻滞,双基因联合使HepG2细胞同时发生G2/M期和G0/G1期阻滞,是IL-24和DCN对HepG2细胞增殖抑制和凋亡诱导协同效应的可能机制.
目的 觀察外源基因重組人源白細胞介素(IL)-24(rhIL-24)聯閤重組人源覈心蛋白聚糖(rhDCN)對人類肝細胞癌HepG2細胞的增殖、凋亡和細胞週期的影響.方法 採用脂質體瞬時轉染法將pcDNA3.1(+)-IL-24和pcDNA3.1(+)-DCN質粒轉染至HepG2細胞,48 h後倒置顯微鏡下觀察細胞生長狀況、形態學改變.分彆在轉染後24h、48 h和72 h,採用四甲基偶氮唑藍(MTT)法觀察IL-24和DCN單獨及聯閤轉染對HepG2細胞的增殖抑製效應.轉染後48 h流式細胞術檢測HepG2細胞凋亡情況,併進行細胞週期阻滯分析.結果 與對照組相比,轉染目的基因48 h後顯微鏡下可見細胞生長不同程度地被抑製,併可見典型的凋亡細胞形態改變,以聯閤轉染組改變更為明顯.MTT結果顯示轉染後48 h和72 h,雙基因聯閤轉染組的抑製率分彆是(31.88±6.57)％和(36.83±3.76)％,與對照組和單獨轉染組比較,差異均有統計學意義(P＜0.01).流式細胞術結果顯示,單獨轉染IL-24和DCN基因可誘導HepG2細胞齣現不同程度凋亡,而雙基因聯閤轉染組細胞凋亡率可達(32.56±0.90)％,與空質粒轉染組的(2.98±0.72)％、空白細胞對照組的(3.50±0.92)％、IL-24組的(20.01±1.08)％和DCN組的(22.20±0.91)％比較,差異均有統計學意義(P＜0.01).細胞週期分析結果錶明,與對照組相比,IL-24基因轉染組的G2/M期和DCN基因轉染組的G0/G1期細胞比例明顯增高分彆為(11.24±0.35)％、(77.93±0.67)％,而雙基因聯閤轉染組G0/G1期和G2/M期細胞比例同時增高,分彆是(71.36±0.60)％和(10.39±0.67)％,差異有統計學意義(P＜0.01).結論 聯閤轉染IL-24和DCN基因可以協同髮揮較單獨應用更彊的抑製HepG2細胞增殖和誘導HepG2細胞凋亡的作用.IL-24可使HepG2細胞髮生G2/M期阻滯,DCN可使HepG2細胞髮生G0/G1期阻滯,雙基因聯閤使HepG2細胞同時髮生G2/M期和G0/G1期阻滯,是IL-24和DCN對HepG2細胞增殖抑製和凋亡誘導協同效應的可能機製.
목적 관찰외원기인중조인원백세포개소(IL)-24(rhIL-24)연합중조인원핵심단백취당(rhDCN)대인류간세포암HepG2세포적증식、조망화세포주기적영향.방법 채용지질체순시전염법장pcDNA3.1(+)-IL-24화pcDNA3.1(+)-DCN질립전염지HepG2세포,48 h후도치현미경하관찰세포생장상황、형태학개변.분별재전염후24h、48 h화72 h,채용사갑기우담서람(MTT)법관찰IL-24화DCN단독급연합전염대HepG2세포적증식억제효응.전염후48 h류식세포술검측HepG2세포조망정황,병진행세포주기조체분석.결과 여대조조상비,전염목적기인48 h후현미경하가견세포생장불동정도지피억제,병가견전형적조망세포형태개변,이연합전염조개변경위명현.MTT결과현시전염후48 h화72 h,쌍기인연합전염조적억제솔분별시(31.88±6.57)％화(36.83±3.76)％,여대조조화단독전염조비교,차이균유통계학의의(P＜0.01).류식세포술결과현시,단독전염IL-24화DCN기인가유도HepG2세포출현불동정도조망,이쌍기인연합전염조세포조망솔가체(32.56±0.90)％,여공질립전염조적(2.98±0.72)％、공백세포대조조적(3.50±0.92)％、IL-24조적(20.01±1.08)％화DCN조적(22.20±0.91)％비교,차이균유통계학의의(P＜0.01).세포주기분석결과표명,여대조조상비,IL-24기인전염조적G2/M기화DCN기인전염조적G0/G1기세포비례명현증고분별위(11.24±0.35)％、(77.93±0.67)％,이쌍기인연합전염조G0/G1기화G2/M기세포비례동시증고,분별시(71.36±0.60)％화(10.39±0.67)％,차이유통계학의의(P＜0.01).결론 연합전염IL-24화DCN기인가이협동발휘교단독응용경강적억제HepG2세포증식화유도HepG2세포조망적작용.IL-24가사HepG2세포발생G2/M기조체,DCN가사HepG2세포발생G0/G1기조체,쌍기인연합사HepG2세포동시발생G2/M기화G0/G1기조체,시IL-24화DCN대HepG2세포증식억제화조망유도협동효응적가능궤제.
Objective To investigate the synergistic proliferation-inhibiting and apoptosis-inducing effects of recombinant human-derived interleukin-24 (rhIL-24) and recombinant human-derived decorin (rhDCN) on human hepatocellular carcinoma cells HepG2.Methods Cellular growth and morphological changes of HepG2 cells were observed under the inverted microscope at 48 h after being transiently transfected with pcDNA3.1 (+)-IL-24 and pcDNA3.1 (+)-DCN by Lipofectamine.The proliferation-inhibiting effects of IL-24 and DCN on HepG2 cells,respectively and jointly,were observed with MTT assay at 24 h,48 h and 72 h post-transfection.Apoptosis and cell-cycle of HepG2 cells were analyzed by flow cytometry at 48 h post-transfection.Results Compared to control groups,the cells of target gene groups presented typically changes of proliferation inhibition and apoptotic morphology,which occurred obviously in co-transfection group.The results of MTT assay showed that at 48 h and 72 h post-transfection,the profiferation-inhibiting rates in the group of cells co-transfected with IL-24 and DCN were (31.88±6.57) ％ and (36.83±3.76) ％,respectively,displaying significant difference with those of other groups (P ＜ 0.01).The results of flow cytometry showed that IL-24 and DCN can induce HepG2 cells apoptosis to some extent.Compared to the early apoptosis rate of cells of control groups,plasmid (2.98±0.72) ％,blank cell (3.50±0.92) ％,IL-24 (20.01±1.08) ％ and DCN (22.20±0.91) ％,a statistically remarkable apoptosis rate,(32.56±0.90) ％,can be seen in the group of cells treated with IL-24 and DCN jointly (P ＜ 0.01).The result of cell cycle analysis revealed that,compared to control groups,the proportion of cells was higher in the phase of G2/M in the IL-24 group (11.24±0.35) ％ and in the phase of G0/G1 in the DCN group (77.93±0.67) ％.The proportions of cells in the phases of G2/M increased to (71.36±0.60) ％ and that of G0/G1 statistically increased to (10.39±0.67) ％ in the group of cells co-transfected with IL-24 and DCN (P ＜ 0.01).Conclusions Combinatorial treatment of HepG2 cells with IL-24 and DCN can exert stronger synergistic proliferation-inhibiting effect and apoptosisinducing activity-in comparison to single therapies.IL-24 and DCN can induce cell cycle arrest on HepG2 cells,occurred in the phase of G2/M and G0/G1,respectively.Promoting effect of cell cycle arrest in the phase of both G2/M and G0/G1 can be seen on HepG2 cells co-transfected with IL-24 and DCN,which maybe the possible mechanism of the synergistic proliferation-inhibiting and apoptosis-inducing effect.
