癌变·畸变·突变
癌變·畸變·突變
암변·기변·돌변
CARCINOGENSES,TERATOGENSIS AND MUTAGENESIS
2015年
3期
172-176
,共5页
梁斌%袁松%刘艳敏%宋旭红
樑斌%袁鬆%劉豔敏%宋旭紅
량빈%원송%류염민%송욱홍
腺苷酸活化蛋白激酶%肿瘤细胞%内质网%应激
腺苷痠活化蛋白激酶%腫瘤細胞%內質網%應激
선감산활화단백격매%종류세포%내질망%응격
AMPK%cancer cells%endoplasmic reticulum%stress
目的：探讨腺苷酸活化蛋白激酶(AMPK)对HeLa细胞内质网应激的影响。方法：试验分为对照组(1‰ DMSO)、内质网应激诱导组(2μg/mL衣霉素或500 nmol/L MG132内质网应激诱导剂诱导)和内质网应激干预组[衣霉素或MG132+2 mmol/L AMPK激动剂5-氨基咪唑-4-甲酰胺-1-B-呋喃核糖苷(AICAR)干预组、衣霉素或MG132+2 mmol/L AMPK激动剂二甲双胍干预组以及衣霉素或MG132+0.5 mmol/L AMPK抑制剂Compound C干预组]，以探究各处理组中AMPK活性状态对内质网应激的影响。收集对数生长期的HeLa细胞，分组诱导孵育后，用Western blot检测HeLa细胞AMPK/T-172磷酸化水平及内质网应激标志蛋白(p-elf2α、Grp78和XBP-2+2+1s)的表达，并采用细胞内钙集群检测分析(Indo-1 ratiometric Ca analysis)检测各组HeLa细胞内Ca浓度的变化。结果：衣霉素和MG132可诱导内质网应激标志蛋白p-elf2α、Grp78和XBP-1s的表达。AMPK激动剂AICAR和二甲双胍干预可降低上述3种内质网应2+激标志蛋白的表达，并降低细胞胞浆内因内质网应激导致的Ca 浓度升高，与对照组比较，差异均有统计学意义(P均<0.05)。AMPK抑制剂Compound C的干预对上述内质网应激标志蛋白的表达无明显影响(P均>0.05)。结论：AMPK激活能缓解HeLa细胞中的内质网应激，维持细胞内质网的功能稳态，可能对提高肿瘤细胞的应激耐受能力起到主要作用。
目的：探討腺苷痠活化蛋白激酶(AMPK)對HeLa細胞內質網應激的影響。方法：試驗分為對照組(1‰ DMSO)、內質網應激誘導組(2μg/mL衣黴素或500 nmol/L MG132內質網應激誘導劑誘導)和內質網應激榦預組[衣黴素或MG132+2 mmol/L AMPK激動劑5-氨基咪唑-4-甲酰胺-1-B-呋喃覈糖苷(AICAR)榦預組、衣黴素或MG132+2 mmol/L AMPK激動劑二甲雙胍榦預組以及衣黴素或MG132+0.5 mmol/L AMPK抑製劑Compound C榦預組]，以探究各處理組中AMPK活性狀態對內質網應激的影響。收集對數生長期的HeLa細胞，分組誘導孵育後，用Western blot檢測HeLa細胞AMPK/T-172燐痠化水平及內質網應激標誌蛋白(p-elf2α、Grp78和XBP-2+2+1s)的錶達，併採用細胞內鈣集群檢測分析(Indo-1 ratiometric Ca analysis)檢測各組HeLa細胞內Ca濃度的變化。結果：衣黴素和MG132可誘導內質網應激標誌蛋白p-elf2α、Grp78和XBP-1s的錶達。AMPK激動劑AICAR和二甲雙胍榦預可降低上述3種內質網應2+激標誌蛋白的錶達，併降低細胞胞漿內因內質網應激導緻的Ca 濃度升高，與對照組比較，差異均有統計學意義(P均<0.05)。AMPK抑製劑Compound C的榦預對上述內質網應激標誌蛋白的錶達無明顯影響(P均>0.05)。結論：AMPK激活能緩解HeLa細胞中的內質網應激，維持細胞內質網的功能穩態，可能對提高腫瘤細胞的應激耐受能力起到主要作用。
목적：탐토선감산활화단백격매(AMPK)대HeLa세포내질망응격적영향。방법：시험분위대조조(1‰ DMSO)、내질망응격유도조(2μg/mL의매소혹500 nmol/L MG132내질망응격유도제유도)화내질망응격간예조[의매소혹MG132+2 mmol/L AMPK격동제5-안기미서-4-갑선알-1-B-부남핵당감(AICAR)간예조、의매소혹MG132+2 mmol/L AMPK격동제이갑쌍고간예조이급의매소혹MG132+0.5 mmol/L AMPK억제제Compound C간예조]，이탐구각처리조중AMPK활성상태대내질망응격적영향。수집대수생장기적HeLa세포，분조유도부육후，용Western blot검측HeLa세포AMPK/T-172린산화수평급내질망응격표지단백(p-elf2α、Grp78화XBP-2+2+1s)적표체，병채용세포내개집군검측분석(Indo-1 ratiometric Ca analysis)검측각조HeLa세포내Ca농도적변화。결과：의매소화MG132가유도내질망응격표지단백p-elf2α、Grp78화XBP-1s적표체。AMPK격동제AICAR화이갑쌍고간예가강저상술3충내질망응2+격표지단백적표체，병강저세포포장내인내질망응격도치적Ca 농도승고，여대조조비교，차이균유통계학의의(P균<0.05)。AMPK억제제Compound C적간예대상술내질망응격표지단백적표체무명현영향(P균>0.05)。결론：AMPK격활능완해HeLa세포중적내질망응격，유지세포내질망적공능은태，가능대제고종류세포적응격내수능력기도주요작용。
OBJECTIVE:To observe the effects of AMPK activation on intracellular endoplasmic reticulum stress (ER stress) in Hela cells.METHODS:Cultured HeLa cells were divided into groups:1‰ DMSO control,tunicamycin (2μg/mL) or MG132 (500 nmol/L),tunicamycin or MG132+Compound C (0.5 mmol/L);tunicamycin or MG132+ AICAR(2 mmol/L) and tunicamycin or MG132+metformin (2 mmol/L). AMPK activation and markers of ER stress were 2+determined by Western blot,and then intracellular Ca concentration was measured in HeLa cells by Indo-1 ratiometric 2+Ca analysis.RESULTS:Our results showed that tunicamycin and MG132 could increase expression of ER stress. Activation of AMPK by AICAR and metformin could reduce the expression of above markers,and decrease the 2+concentration of intracellular Ca.CONCLUSION:AMPK activation could relieve intracellular ER stress,maintain the 2+homeostasis of intracellular Ca concentration,and further,may contribute to cellular adaptation and resistance to stress stimuli.
