熱CO2誘導(dǎo)胃癌細(xì)胞凋亡并抑制其侵襲遷移能力的研究.pdf

1、張繼燃熱CO!誘導(dǎo)胃癌細(xì)胞凋亡并抑制其侵襲遷移能力的研究熱C02誘導(dǎo)胃癌細(xì)胞凋亡并抑制其侵襲遷移能力的研究摘要目的：體外模擬熱C02氣腹環(huán)境，通過細(xì)胞實(shí)驗(yàn)，初步探討熱C02對(duì)胃癌細(xì)胞SGC一7901增殖、凋亡及侵襲遷移能力的影響，由此可為腹腔鏡胃癌手術(shù)時(shí)應(yīng)用熱C02氣腹的可行性和安全性奠定一定的理論基礎(chǔ)。方法：建立熱C02氣腹體外實(shí)驗(yàn)?zāi)Ｐ?，由C02鋼瓶、C02加熱器及氣腹箱組成：胃癌細(xì)胞SGC一7901分組處理：對(duì)照組(細(xì)胞培養(yǎng)箱中培養(yǎng)

2、)，單純熱療組(42。C環(huán)境中培養(yǎng)3h)，單純C02組(注入常溫C02，培養(yǎng)細(xì)胞3h)；熱C02組(注入429CC02，培養(yǎng)細(xì)胞3h)。CCK一8法檢測(cè)各組細(xì)胞增殖情況，AnnexinV—FITC／PI流式細(xì)胞術(shù)及Hoechst33342／PI雙染熒光顯微技術(shù)檢測(cè)細(xì)胞凋亡情況，透射電鏡觀察各組細(xì)胞形態(tài)學(xué)改變，細(xì)胞劃痕實(shí)驗(yàn)及transwell小室遷移實(shí)驗(yàn)檢測(cè)細(xì)胞侵襲遷移能力的改變。實(shí)驗(yàn)數(shù)據(jù)由均數(shù)土標(biāo)準(zhǔn)差(X士s)表示，通過SPSSl70軟

3、件分析，p005差異有統(tǒng)計(jì)學(xué)意義。結(jié)果：CCK8法檢測(cè)細(xì)胞增殖情況，結(jié)果顯示：對(duì)照組增殖抑制率為0，單純C02組細(xì)胞增殖抑制率為1176i729％，單純熱療組細(xì)胞增殖抑制率為2808士677％，熱C02組細(xì)胞增殖抑制率為3893士763％，通過統(tǒng)計(jì)分析，差異有統(tǒng)計(jì)學(xué)意義，熱C02氣腹能夠顯著抑制胃癌細(xì)胞增殖p0。05)；AnnexinV—FITC／PI流式細(xì)胞術(shù)檢測(cè)結(jié)果顯示：對(duì)照組，單純C02組，單純熱療組，熱C02組細(xì)胞凋亡率分別為0

4、033士0015％，264士021％，350土022％，491土O20％，差異有統(tǒng)計(jì)學(xué)意義，熱C02顯著誘導(dǎo)胃癌細(xì)胞凋亡(p005)；Hoechst33342／P1雙染熒光顯微鏡技術(shù)檢測(cè)結(jié)果顯示：與其余3組細(xì)胞相比，熱C02組細(xì)胞出現(xiàn)更多被染成亮藍(lán)色或亮紅色的凋亡壞死細(xì)胞，由此說明，熱C02組細(xì)胞發(fā)生了顯著凋亡：通過透射電鏡進(jìn)行細(xì)胞形態(tài)學(xué)檢測(cè)結(jié)果顯示：熱C02組細(xì)胞的細(xì)胞質(zhì)濃縮，染色質(zhì)固縮、邊集，核碎裂，凋亡小體形成，其細(xì)胞膜及細(xì)胞核發(fā)

5、生凋亡形態(tài)學(xué)變化；細(xì)胞劃痕張繼燃熱C02誘導(dǎo)胃癌細(xì)胞凋亡并抑制其侵襲遷移能力的研究InducedapoptosisandinhibitedcapabilityofinvasionandmigrationforgastriccancercellsbyhyperthermicC02Abstract3Objectives：WepreliminaryinvestigatedtheimpactofahyperthermicC02pneumoper

6、itoneumongastriccancercellsonaspectofproliferation，apoptosis，invasiveandmigrateabilitybycellexperimentofinvitrohyperthermicC02pneumoperitoneumcircumstancesThiscouldformthetheoreticalbasisforfurtherstudiesofthefeasibility

7、andsafetyofinflatinghyperthermicC02intheabdominalcavityofgastriccancerpatientsduring1aparoscopyMethods：AninvitrohyperthermicC02pneumoperitoneumexperimentalmodelwasbuilt，whichwascomposedofC02steel，C02heaterandpneumoperito

8、neumboxGastriccancercelllineSGC一7901cellsweregroupedaccordingtotemperature：controlgroup(conventionalcultureinincubator)，purethermotherapygroup(culturedin42。Cfor3h)，pureC02group(culturedinc02environmentofnormaltemperature

9、for3h)，hyperthermicC02groupfculturedinC02environmentof42℃for3h)Cellproliferationwasdetectedusingacellcountingkit一8(CCK一8)；apoptosiswasdetectedbyAnnexinV—FITC／PItlOWcytometryandHoechst33342／PIfluorescentmicroscopyMorpholo

10、gicalalterationswereobservedbytransmissionelectronmicroscopyInvasionandmigrationweredetectedbYascratchtestandbytranswellmigration，respectivelyTheexperimentaldatawasanalyzedbySPSS170andshownbythearithmeticmeanthestandardd

11、eviation(X土s)APvalueO05indicatedastatisticallysignificantdifferenceResults：CCK一8wasusedtodetecttheinhibitionofcellproliferation，theresultsshOW；proliferationofthepureC02groupwasinhibitedby1176士729％，ofthepurethermotherapyg