本文選題：硼替佐米 + 蛋白酶體抑制劑�。� 參考：《蘇州大學(xué)》2014年碩士論文

【摘要】：目的：宮頸癌為大家所熟知,也是目前所知的婦科最為常見的惡性腫瘤,每年全球都約有二十幾萬的婦女以為此而去世,宮頸癌對婦女的身心健康和生存品質(zhì)都有著嚴(yán)重的負(fù)面影響,給許多家庭帶來了巨大的經(jīng)濟(jì)負(fù)擔(dān)。宮頸癌一般的治療手段主要是以手術(shù)和放化療,這些手段可以使一部分病患得到根治或著相對的緩解病情,但大多數(shù)的病患經(jīng)過手術(shù)或者放化療措施后的遠(yuǎn)期預(yù)后并非特別理想,且部分人群對化療又不是很敏感,甚至產(chǎn)生抵抗,故預(yù)后不良。因此為了全人類的健康及生存環(huán)境,迫切需要尋找對宮頸癌較為有效的方法從而大大改善宮頸癌患者的預(yù)后。硼替佐米(Bortezomib)是一種細(xì)胞內(nèi)蛋白酶體抑制劑的抑制劑,人體細(xì)胞內(nèi)的蛋白酶體的糜蛋白酶樣活性和胰蛋白酶樣活性這兩者都可以受到Bortezomib的抑制作用。5-FU脫氧核苷酸,5-FU核苷等代謝產(chǎn)物可以由5-氟尿嘧啶(5-FU)在人體細(xì)胞內(nèi)轉(zhuǎn)化而成,這些物質(zhì)由于和DNA及蛋白質(zhì)的合成所需底物相似,從而影響或干擾細(xì)胞內(nèi)正常的代謝過程,故5-FU作為一種通過影響細(xì)胞代謝而抗腫瘤的藥物,在臨床惡性腫瘤的治療方面已被廣泛地采用,本文主要探討的是Bortezomib這一血液腫瘤常用的化療藥物和常見的臨床腫瘤化療藥物5-氟尿嘧啶(5-FU)對于Hela細(xì)胞的作用是怎樣的,并淺表分析其作用機(jī)制。 方法：將人的宮頸腺癌Hela細(xì)胞培養(yǎng)于培養(yǎng)瓶中,呈單層貼壁樣生長,培養(yǎng)基為含10%新生牛血清的DMEM-H(高糖)完全培養(yǎng)基,并將培養(yǎng)瓶放置于擁有飽和的濕度、溫度為37攝氏度、含5%二氧化碳環(huán)境適合人類細(xì)胞生存的培養(yǎng)箱中進(jìn)行傳代培養(yǎng)。分為空白對照組,單純Bortezomib組,單純5-FU組及聯(lián)合組(Bortezomib和5-FU聯(lián)合應(yīng)用組)4大組。并通過MTT法來測定各個組別的細(xì)胞抑制率情況。采用RT-PCR法測定不同組別中hela細(xì)胞的Bcl-2、Survivn、IGF-1及HIF-lamRNA的表達(dá)；PI標(biāo)記流式細(xì)胞術(shù)(FCM)分析不同組別的細(xì)胞周期、AnnexinV/PI雙染法法測定不同組別的凋亡的變化。 結(jié)果：(1)MTT檢測法的檢測結(jié)果顯示,Bortezomib及5-FU單獨(dú)使用時均可抑制Hela細(xì)胞的增殖,且抑制作用與作用的時間及濃度呈正相關(guān),兩者聯(lián)合作用后抑制增殖作用更顯著(P0.05);(2)RT-PCR結(jié)果顯示Bortezomib及5-FU均可使BCL-2、 IGF mRNA及Survivn mRNA的表達(dá)下降,且與藥物濃度呈正比,并且聯(lián)合化療組較單獨(dú)化療組的抑制作用更為顯著(P0.05); Bortezomib及5-FU對HIF1αmRNA表達(dá)無抑制作用,但兩者聯(lián)合應(yīng)用后其表達(dá)量較對照組顯著降低(P0.05)。(3)細(xì)胞周期結(jié)果顯示Bortezomib及5-FU可影響Hela細(xì)胞的周期比例發(fā)生變化,使其處于S期的細(xì)胞比例減少,聯(lián)合化療組較單獨(dú)化療組更加顯著(P0.05)。(4)凋亡結(jié)果顯示Bortezomib和5-FU單獨(dú)使用時均可引誘導(dǎo)Hela細(xì)胞發(fā)生凋亡,聯(lián)合化療組誘導(dǎo)凋亡的作用較單獨(dú)化療組效果更加顯著(P0.05)。 結(jié)論：本實(shí)驗(yàn)結(jié)果表明,Bortezomib聯(lián)合5-FU應(yīng)用可以明顯的增加人宮頸腺癌Hela細(xì)胞的凋亡率,而降低Hela細(xì)胞的增殖率。
[Abstract]:Objective: cervical cancer is well known and is also known as the most common malignant tumor of gynecology. Every year, more than 20 million women all over the world have died. Cervical cancer has a serious negative impact on the physical and mental health and quality of life of women. It has brought great economic burden to many families. The general treatment of cervical cancer. The treatment means mainly by operation and radiotherapy and chemotherapy, these methods can make a part of the patients get a radical cure or relative relief of the condition, but the long-term prognosis of most patients after surgery or chemotherapy is not particularly ideal, and some people are not very sensitive to chemotherapy and even produce resistance, so the prognosis is bad. The health and living environment of all human beings urgently need to find a more effective method for cervical cancer to greatly improve the prognosis of cervical cancer patients. Borteg Zomi (Bortezomib) is an inhibitor of intracellular proteasome inhibitors, both the chymotrypsin like activity and trypsin like activity of the proteasome in the human body The metabolites such as.5-FU deoxynucleotides, 5-FU nucleosides, and 5-FU nucleosides can be transformed by 5- fluorouracil (5-FU) in human cells, which affect or interfere with normal metabolic processes in cells due to the similar substrates required for the synthesis of DNA and protein, so 5-FU acts as a way of affecting cell metabolism. Antitumor drugs have been widely used in the treatment of clinical malignant tumors. This article mainly discusses how the chemotherapeutic drugs commonly used in the Bortezomib blood tumor and the common clinical tumor chemotherapeutic drugs, 5- fluorouracil (5-FU), have the effect on the Hela cells.
Methods: human cervical adenocarcinoma Hela cells were cultured in a culture bottle with monolayer adherent growth. The culture medium was a DMEM-H (high sugar) complete medium containing 10% newborn bovine serum, and the culture bottle was placed in the incubator with saturated humidity, temperature of 37 degrees Celsius, and 5% carbon dioxide in the incubator suitable for human cell survival. It was divided into 4 groups: blank control group, simple Bortezomib group, simple 5-FU group and combined group (Bortezomib and 5-FU combined application group). The cell inhibition rate of each group was measured by MTT method. The expression of Bcl-2, Survivn, IGF-1 and HIF-lamRNA of HeLa cells in different groups was measured by RT-PCR; PI labeled flow cytometry was divided into two groups. The cell cycle of different groups was analyzed, and the changes of apoptosis in different groups were determined by AnnexinV/PI double staining.
Results: (1) the results of MTT detection showed that the proliferation of Hela cells could be inhibited when Bortezomib and 5-FU were used alone, and the inhibition effect was positively correlated with the time and concentration of the action, and the inhibition of proliferation was more significant (P0.05). (2) RT-PCR knots showed that Bortezomib and 5-FU could make BCL-2, IGF mRNA and Survivn. The expression decreased and was proportional to the drug concentration, and the inhibitory effect of the combined chemotherapy group was more significant than that of the chemotherapy group (P0.05); Bortezomib and 5-FU had no inhibitory effect on the expression of HIF1 alpha mRNA, but the expression of the two groups was significantly lower than that of the control group (P0.05). (3) the cell cycle results showed that Bortezomib and 5-FU could affect Hela fine. The proportion of cell cycle was changed, the proportion of cells in S phase decreased, and the combined chemotherapy group was more significant than that in the chemotherapy group (P0.05). (4) apoptosis results showed that Bortezomib and 5-FU could induce apoptosis of Hela cells, and the effect of combined chemotherapy group to induce apoptosis was more significant than that of chemotherapy group (P0.05).
Conclusion: the results show that Bortezomib combined with 5-FU can significantly increase the apoptosis rate of human cervical adenocarcinoma Hela cells, and decrease the proliferation rate of Hela cells.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.33
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本文編號：2028702