中华生物医学工程杂志
中華生物醫學工程雜誌
중화생물의학공정잡지
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
2009年
5期
354-359
,共6页
廖洪映%谷力加%翁毅敏%黄邵洪%陈惠国%李昀%蔡松旺%张健%宋江平
廖洪映%穀力加%翁毅敏%黃邵洪%陳惠國%李昀%蔡鬆旺%張健%宋江平
료홍영%곡력가%옹의민%황소홍%진혜국%리윤%채송왕%장건%송강평
RNA干扰%缺氧诱导因子1%α亚基%顺铂%食管肿瘤%鳞癌
RNA榦擾%缺氧誘導因子1%α亞基%順鉑%食管腫瘤%鱗癌
RNA간우%결양유도인자1%α아기%순박%식관종류%린암
RNA interference%Hypoxia-inducible factor 1%alpha subunit%Cisplatin%Esophageal neoplasm%squamous carcinoma
目的 探讨缺氧诱导因子1α(HIF-1α)小干扰RNA(siRNA)联合顺铂对人食管鳞癌TE-1细胞裸鼠移植瘤生长的影响.方法 体外培养人食管鳞癌TE-1细胞,接种于12只裸鼠右上肢外侧皮下,建立裸鼠荷瘤模型开始抑瘤实验.实验分为4组,每组3只,A组用生理盐水瘤内注射;B组用顺铂(20mg/L)瘤内注射;C组用HIF-1α siRNA(20 μmol/L)瘤内注射;D组用HIF-1α siRNA(20 μmol/L)和顺铂(20 mg/L)协同瘤内注射.各组注射体积均为20 μl,每周测量肿瘤体积变化.3周后处死裸鼠,测量各组移植瘤质量;RT-PCR半定量检测各组移植瘤TE-1细胞HIF-1α mRNA的表达;免疫组织化学法检测其HIF-1α蛋白的表达;流式细胞仪AnnexinV-FITC/PI双标检测其细胞的凋亡.结果 抑瘤实验开始3周后,A、B、C和D组的移植瘤体积分别为(1.477±0.132)cm~3、(1.200±0.114)cm~3、(1.223±0.129)cm~3 和(0.890±0.141)cm~3;移植瘤质量分别为(7.38±0.96)g、(6.35±0.73)g、(6.12±0.65)g和(3.51±0.42)g.B、C、D 3组的抑瘤率分别为14.0%、17.1%、52.4%,其中D组移植瘤的生长受抑制最明显,与其他3组比较差异有统计学意义(P<0.05).C和D组TE-1细胞的HIF-1α mRNA表达下调至0.343±0.080和0.312±0.055,蛋白表达下调至0.196±0.018和0.229±0.035,与A(mRNA 1.315±0.179,蛋白0.523±0.102)、B(mRNA 1.483±0.460,蛋白0.542±0.174)两组差异有统计学意义(P<0.01),而C、D两组间差异无统计学意义(P>0.05).A、B、C和D组的TE-1细胞凋亡率分别为(6.13±1.38)%、(28.30±4.82)%、(34.10+5.56)%、(52.88±8.77)%,其中D组与其他3组比较,差异有统计学意义(P<0.01).结论 HIF-1α siRNA能有效下调裸鼠体内食管鳞癌TE-1细胞HIF-1α的表达,提高顺铂的细胞毒性作用,诱导细胞凋亡,显著抑制移植瘤的生长.
目的 探討缺氧誘導因子1α(HIF-1α)小榦擾RNA(siRNA)聯閤順鉑對人食管鱗癌TE-1細胞裸鼠移植瘤生長的影響.方法 體外培養人食管鱗癌TE-1細胞,接種于12隻裸鼠右上肢外側皮下,建立裸鼠荷瘤模型開始抑瘤實驗.實驗分為4組,每組3隻,A組用生理鹽水瘤內註射;B組用順鉑(20mg/L)瘤內註射;C組用HIF-1α siRNA(20 μmol/L)瘤內註射;D組用HIF-1α siRNA(20 μmol/L)和順鉑(20 mg/L)協同瘤內註射.各組註射體積均為20 μl,每週測量腫瘤體積變化.3週後處死裸鼠,測量各組移植瘤質量;RT-PCR半定量檢測各組移植瘤TE-1細胞HIF-1α mRNA的錶達;免疫組織化學法檢測其HIF-1α蛋白的錶達;流式細胞儀AnnexinV-FITC/PI雙標檢測其細胞的凋亡.結果 抑瘤實驗開始3週後,A、B、C和D組的移植瘤體積分彆為(1.477±0.132)cm~3、(1.200±0.114)cm~3、(1.223±0.129)cm~3 和(0.890±0.141)cm~3;移植瘤質量分彆為(7.38±0.96)g、(6.35±0.73)g、(6.12±0.65)g和(3.51±0.42)g.B、C、D 3組的抑瘤率分彆為14.0%、17.1%、52.4%,其中D組移植瘤的生長受抑製最明顯,與其他3組比較差異有統計學意義(P<0.05).C和D組TE-1細胞的HIF-1α mRNA錶達下調至0.343±0.080和0.312±0.055,蛋白錶達下調至0.196±0.018和0.229±0.035,與A(mRNA 1.315±0.179,蛋白0.523±0.102)、B(mRNA 1.483±0.460,蛋白0.542±0.174)兩組差異有統計學意義(P<0.01),而C、D兩組間差異無統計學意義(P>0.05).A、B、C和D組的TE-1細胞凋亡率分彆為(6.13±1.38)%、(28.30±4.82)%、(34.10+5.56)%、(52.88±8.77)%,其中D組與其他3組比較,差異有統計學意義(P<0.01).結論 HIF-1α siRNA能有效下調裸鼠體內食管鱗癌TE-1細胞HIF-1α的錶達,提高順鉑的細胞毒性作用,誘導細胞凋亡,顯著抑製移植瘤的生長.
목적 탐토결양유도인자1α(HIF-1α)소간우RNA(siRNA)연합순박대인식관린암TE-1세포라서이식류생장적영향.방법 체외배양인식관린암TE-1세포,접충우12지라서우상지외측피하,건립라서하류모형개시억류실험.실험분위4조,매조3지,A조용생리염수류내주사;B조용순박(20mg/L)류내주사;C조용HIF-1α siRNA(20 μmol/L)류내주사;D조용HIF-1α siRNA(20 μmol/L)화순박(20 mg/L)협동류내주사.각조주사체적균위20 μl,매주측량종류체적변화.3주후처사라서,측량각조이식류질량;RT-PCR반정량검측각조이식류TE-1세포HIF-1α mRNA적표체;면역조직화학법검측기HIF-1α단백적표체;류식세포의AnnexinV-FITC/PI쌍표검측기세포적조망.결과 억류실험개시3주후,A、B、C화D조적이식류체적분별위(1.477±0.132)cm~3、(1.200±0.114)cm~3、(1.223±0.129)cm~3 화(0.890±0.141)cm~3;이식류질량분별위(7.38±0.96)g、(6.35±0.73)g、(6.12±0.65)g화(3.51±0.42)g.B、C、D 3조적억류솔분별위14.0%、17.1%、52.4%,기중D조이식류적생장수억제최명현,여기타3조비교차이유통계학의의(P<0.05).C화D조TE-1세포적HIF-1α mRNA표체하조지0.343±0.080화0.312±0.055,단백표체하조지0.196±0.018화0.229±0.035,여A(mRNA 1.315±0.179,단백0.523±0.102)、B(mRNA 1.483±0.460,단백0.542±0.174)량조차이유통계학의의(P<0.01),이C、D량조간차이무통계학의의(P>0.05).A、B、C화D조적TE-1세포조망솔분별위(6.13±1.38)%、(28.30±4.82)%、(34.10+5.56)%、(52.88±8.77)%,기중D조여기타3조비교,차이유통계학의의(P<0.01).결론 HIF-1α siRNA능유효하조라서체내식관린암TE-1세포HIF-1α적표체,제고순박적세포독성작용,유도세포조망,현저억제이식류적생장.
Objective To study the effect of hypoxia-inducible factor la (HIF-1α) small interfering RNA (siRNA) combined with cisplatin on growth of human esophageal squamous carcinoma cells transplanted in nude mice. Methods Human esophageal squamous carcinoma TE-1 cells cultured in vitro were transplanted under the lateral skin of right upper limb in 12 nude mice to establish tumor-bearing models. Anti-tumor experiment was carried out. The nude mice were then divided into four groups (n=3 for each) to receive intratumor injection of normal saline (Group A), 20 mg/L cisplatin (Group B), 20 μmol/L HIF-1α siRNA (Group C) or both 20 mg/L cisplatin and 20 μmol/L HIF-1α siRNA (Group D). The volume of injected agent was 20 μl for each group. Change in tumor size was recorded every week. At week 3, all the nude mice were sacrificed and the transplanted tumor volume and weight were measured. In transplanted tumor of all the mice, the expression of HIF-1 α mRNA was detected by semi-quantitative RT-PCR and HIF-1 α protein was detected by immunohistochemistry. The apoptosis of TE-1 cells was determined by flow cytometry with Annexin V-FITC/PI dual staining. Results After anti-tumor experiment for three weeks, the transplanted tumor volumes of groups A,B,C and D were (1.477±0.132) cm~3, (1.200±0.114) cm~3, (1.223±0.129) cm~3 and (0.890±0.141) cm~3, and the tumor weight was (7.38±0.96) g, (6.35±0.73) g, (6.12±0.65) g and (3.51±0.42) g respectively. The rates of tumor suppression were 14.0%, 17.1% and 52.4% in groups B, C and D. Group D mice experienced strongest tumor suppression as compared with the other 3 groups (P<0.05). The expressions of HIF-1α mRNA and protein were down-regulated to 0.343±0,080 and 0.196±0.018 in Group C, 0.312±0.055 and 0.229±0.035 in Group D with no significant difference between these two groups (P>0.05), but were significantly different with those in Group A (mRNA 1.315± 0.179, protein 0.523±0.102) and Group B (mRNA 1.483±0.460, protein 0.542±0.174)(P<0.01). The rates of TE-1 cell apoptosis in groups A, B, C and D were (6.13±1.38)%, (28.30±4.82)%, (34.10±5.56)% and (52.88±8.77)% respectively, presenting a significant difference when comparing Group D with the other three groups (P<0.01). Conclusion HIF-1α siRNA can down-regulate HIF-1α mRNA and protein expressions in nude mice in vivo, induce cell apoptosis, significantly inhibit tumor growth of esophageal squamous carcinoma TE-1 cells, and improve the cytotoxicity of cisplatin chemotherapy.
