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探討APOBEC3B及APOBEC3G對人類肝癌細胞株Hep3B腫瘤抑制...

國立高雄大學運動健康與休閒學系碩士班

探討APOBEC3B及APOBEC3G對人類肝癌細胞株Hep3B腫瘤抑制的影響 = Study on the tumor suppress of APOBEC3B and APOBEC3G in Human Hepatoma Cell Line,Hep3B

紀錄類型:	書目-語言資料,印刷品 : 單行本
並列題名:	Study on the tumor suppress of APOBEC3B and APOBEC3G in Human Hepatoma Cell Line,Hep3B
作者:	郭婷茵,
其他團體作者:	國立高雄大學
出版地:	[高雄市]
出版者:	撰者;
出版年:	2013[民102]
面頁冊數:	60面圖，表格 : 30公分;
標題:	APOBEC3B
標題:	APOBEC3B
電子資源:	http://handle.ncl.edu.tw/11296/ndltd/73605962792712965161
附註:	參考書目：面50-55
附註:	102年10月31日公開
摘要註:	此研究首先將 APOBEC3B (apolipoprotein B mRNA-editing catalytic polypeptide- like 3B) 及 APOBEC3G (apolipoprotein B mRNA-editing catalytic polypeptide- like 3G) 基因轉移至 pLKO_AS2.puro vector (pLV) 載體中，並轉染入人類肝癌細胞株 Hep3B ，轉染後使用抗體接有 FITC 染色並用螢光顯微鏡觀察，確認 pLV-APOBEC3B 及 pLV-APOBEC3G 在細胞中過度表現的位置，實驗結果顯示 pLV-APOBEC3B 過度表現位置位於細胞核，而 pLV-APOBEC3G 過度表現位置位於細胞質中。為了評估 APOBEC3B 及 APOBEC3G 對於腫瘤抑制的情形，使用 MTS 測試分別轉染 pLV 、 pLV-APOBEC3B 、 pLV-APOBEC3G 的 Hep3B ，轉染後48小時結果發現細胞存活率分別為88%、57%、48%，結果顯示 APOBEC3B 及 APOBEC3G 具有抑制 Hep3B 肝癌細胞株。另外，當 Hep3B 細胞生長至70-80%時，以實驗用微量塑膠吸管尖端於細胞盤上劃一直線，觀察細胞癒合情形發現在72小時，控制組及轉染 pLV 組為100%完全癒合，而分別轉染 pLV-APOBEC3B 及 pLV-APOBEC3G 過度表現的 Hep3B ，細胞癒合程度為48%及43%，結果顯示， APOBEC3B 及 APOBEC3G 具有抑制 Hep3B 細胞生長的能力。最後，轉染 pLV-APOBEC3B 及 pLV-APOBEC3G 於 Hep3B 過度表現後，加入抗腫瘤藥物 Mitomycin-C ， MTS 測試結果顯示， Hep3B 加入3x10-2 mg/ml (90nM) 的 Mitomycin-C 後48小時細胞存活率為49%，結果亦顯示轉染 pLV-APOBEC3B 及 pLV-APOBEC3G 並加入3x10-2 mg/ml的 Mitomycin-C ，細胞存活率分別為27%及32%。綜合實驗結果顯示，APOBEC3B及APOBEC3G具有抑制Hep3B肝癌細胞株的生長能力。 In this study, APOBEC3B (apolipoprotein B mRNA-editing catalytic polypeptide- like 3B) and APOBEC3G (apolipoprotein B mRNA-editing catalytic polypeptide- like 3G) were subcloned into pLKO_AS2.puro vector (pLV) and tranfected to human hepatoma cell line, Hep3B. To clarify where the APOBEC3B and APOBEC3G express locate in cell, FITC stain was apply to the pLV-APOBEC3B and pLV-APOBEC3G overexpressed culture dish and histochemical analysis was observed by fluorescence microscope. Our data shows the overexpressed APOBEC3B was located in cell nucleus and overexpressed APOBEC3G was located in cytoplasma. To evaluate the situation of tumor suppress of APOBEC3B and APOBEC3G in Hep3B, MTS was assayed after pLV , pLV-APOBEC3B and pLV-APOBEC3G were transfected into Hep3B cells. The results show the cell viability was 88%、57% and 48% after pLV-APOBEC3B and pLV-APOBEC3G were transfected into Hep 3B cells for 48hrs, respectively. The results indicate APOBEC3B and APOBEC3G inhibited the cell growth of hepatoma cell line, Hep 3B. Moreover, the Hep 3B cells growth on culture dish were scratched with plastic pipette tip in 70-80% confluence after transfect with pLV-APOBEC3B and pLV-APOBEC3G. Our data shows the ratio of wound closure was 100% in control and pLV express only after wound-scratch 72 hrs observed by phase-contrast microscope. And the ratio of wound migration in pLV-APOBEC3B and pLV-APOBEC3G overexpress was 48% and 43% after wound-scratch 72hrs observed by phase-contrast microscope, respectively. The results also indicated APOBEC3B and APOBEC3G inhibited the cell growth of Hep 3B. Finally, the anti-tumor drug, Mitomycin-C was applied to Hep3B cells after pLV-APOBEC3B and pLV-APOBEC3G were transfected into Hep3B cell. The results of MTS show the cell viability of Hep 3B was 49% after 3x10-2 mg/ml (90nM) Mitomycin-C was applied to Hep 3B cells for 24 hrs. Results also show the cell viability of Hep 3B were 27% and 32% after pLV-APOBEC3B and pLV-APOBEC3G were transfected into Hep 3B cells which containing 3x10-2 mg/ml Mitomycin-C, respectively. Collectively, our results showing here suggest APOBEC3B and APOBEC3G inhibited the cell growth of hepatoma cell line, Hep 3B.

探討APOBEC3B及APOBEC3G對人類肝癌細胞株Hep3B腫瘤抑制的影響 = Study on the tumor suppress of APOBEC3B and APOBEC3G in Human Hepatoma Cell Line,Hep3B
郭, 婷茵

探討APOBEC3B及APOBEC3G對人類肝癌細胞株Hep3B腫瘤抑制的影響 = Study on the tumor suppress of APOBEC3B and APOBEC3G in Human Hepatoma Cell Line,Hep3B / 郭婷茵撰 - [高雄市] : 撰者, 2013[民102]. - 60面 ; 圖，表格 ; 30公分.
參考書目：面50-55102年10月31日公開.
APOBEC3BAPOBEC3B

探討APOBEC3B及APOBEC3G對人類肝癌細胞株Hep3B腫瘤抑制的影響 = Study on the tumor suppress of APOBEC3B and APOBEC3G in Human Hepatoma Cell Line,Hep3B
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328 $a 碩士論文--國立高雄大學運動健康與休閒學系碩士班
330 $a 此研究首先將 APOBEC3B (apolipoprotein B mRNA-editing catalytic polypeptide- like 3B) 及 APOBEC3G (apolipoprotein B mRNA-editing catalytic polypeptide- like 3G) 基因轉移至 pLKO_AS2.puro vector (pLV) 載體中，並轉染入人類肝癌細胞株 Hep3B ，轉染後使用抗體接有 FITC 染色並用螢光顯微鏡觀察，確認 pLV-APOBEC3B 及 pLV-APOBEC3G 在細胞中過度表現的位置，實驗結果顯示 pLV-APOBEC3B 過度表現位置位於細胞核，而 pLV-APOBEC3G 過度表現位置位於細胞質中。為了評估 APOBEC3B 及 APOBEC3G 對於腫瘤抑制的情形，使用 MTS 測試分別轉染 pLV 、 pLV-APOBEC3B 、 pLV-APOBEC3G 的 Hep3B ，轉染後48小時結果發現細胞存活率分別為88%、57%、48%，結果顯示 APOBEC3B 及 APOBEC3G 具有抑制 Hep3B 肝癌細胞株。另外，當 Hep3B 細胞生長至70-80%時，以實驗用微量塑膠吸管尖端於細胞盤上劃一直線，觀察細胞癒合情形發現在72小時，控制組及轉染 pLV 組為100%完全癒合，而分別轉染 pLV-APOBEC3B 及 pLV-APOBEC3G 過度表現的 Hep3B ，細胞癒合程度為48%及43%，結果顯示， APOBEC3B 及 APOBEC3G 具有抑制 Hep3B 細胞生長的能力。最後，轉染 pLV-APOBEC3B 及 pLV-APOBEC3G 於 Hep3B 過度表現後，加入抗腫瘤藥物 Mitomycin-C ， MTS 測試結果顯示， Hep3B 加入3x10-2 mg/ml (90nM) 的 Mitomycin-C 後48小時細胞存活率為49%，結果亦顯示轉染 pLV-APOBEC3B 及 pLV-APOBEC3G 並加入3x10-2 mg/ml的 Mitomycin-C ，細胞存活率分別為27%及32%。綜合實驗結果顯示，APOBEC3B及APOBEC3G具有抑制Hep3B肝癌細胞株的生長能力。 In this study, APOBEC3B (apolipoprotein B mRNA-editing catalytic polypeptide- like 3B) and APOBEC3G (apolipoprotein B mRNA-editing catalytic polypeptide- like 3G) were subcloned into pLKO_AS2.puro vector (pLV) and tranfected to human hepatoma cell line, Hep3B. To clarify where the APOBEC3B and APOBEC3G express locate in cell, FITC stain was apply to the pLV-APOBEC3B and pLV-APOBEC3G overexpressed culture dish and histochemical analysis was observed by fluorescence microscope. Our data shows the overexpressed APOBEC3B was located in cell nucleus and overexpressed APOBEC3G was located in cytoplasma. To evaluate the situation of tumor suppress of APOBEC3B and APOBEC3G in Hep3B, MTS was assayed after pLV , pLV-APOBEC3B and pLV-APOBEC3G were transfected into Hep3B cells. The results show the cell viability was 88%、57% and 48% after pLV-APOBEC3B and pLV-APOBEC3G were transfected into Hep 3B cells for 48hrs, respectively. The results indicate APOBEC3B and APOBEC3G inhibited the cell growth of hepatoma cell line, Hep 3B. Moreover, the Hep 3B cells growth on culture dish were scratched with plastic pipette tip in 70-80% confluence after transfect with pLV-APOBEC3B and pLV-APOBEC3G. Our data shows the ratio of wound closure was 100% in control and pLV express only after wound-scratch 72 hrs observed by phase-contrast microscope. And the ratio of wound migration in pLV-APOBEC3B and pLV-APOBEC3G overexpress was 48% and 43% after wound-scratch 72hrs observed by phase-contrast microscope, respectively. The results also indicated APOBEC3B and APOBEC3G inhibited the cell growth of Hep 3B. Finally, the anti-tumor drug, Mitomycin-C was applied to Hep3B cells after pLV-APOBEC3B and pLV-APOBEC3G were transfected into Hep3B cell. The results of MTS show the cell viability of Hep 3B was 49% after 3x10-2 mg/ml (90nM) Mitomycin-C was applied to Hep 3B cells for 24 hrs. Results also show the cell viability of Hep 3B were 27% and 32% after pLV-APOBEC3B and pLV-APOBEC3G were transfected into Hep 3B cells which containing 3x10-2 mg/ml Mitomycin-C, respectively. Collectively, our results showing here suggest APOBEC3B and APOBEC3G inhibited the cell growth of hepatoma cell line, Hep 3B.
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