發(fā)布時(shí)間：2018-04-26 22:19

  本文選題：宮頸癌 + 治療�。� 參考：《第二軍醫(yī)大學(xué)》2017年碩士論文

【摘要】：宮頸癌,是女性最常見的惡性腫瘤之一,全球每年有近50萬的女性被診斷為宮頸癌,在我國,其發(fā)病率和死亡率占全球近1/4,嚴(yán)重威脅了我國女性的健康。目前認(rèn)為,宮頸癌的發(fā)病與人乳頭瘤病毒(HPV,Human papillomavirus)高危型密切相關(guān)。已有統(tǒng)計(jì)數(shù)據(jù)表明,HPV16,18,31,33,35,39,45,51,52,56,58,59,68,73和82型等均與宮頸癌有關(guān),而其中HPV16型、18型分別在宮頸癌中感染率約為50%和20%;目前大量研究數(shù)據(jù)表明,HPV高危型中的E6及E7蛋白在宮頸癌的惡性維持中起著至關(guān)重要的作用,其中,高危型HPV E6可經(jīng)泛素化途徑誘導(dǎo)腫瘤抑制因子p53的降解,以消除其生長抑制信號(hào)。E7則可磷酸化Rb蛋白,從而發(fā)揮促進(jìn)腫瘤增殖等作用。目前宮頸癌的治療方法主要有手術(shù)及放化療。手術(shù)治療主要適用于宮頸癌早期病人。放療雖然適用于各期病人,但是由于放療可對(duì)陰道及卵巢造成不可逆的損傷,因此,也限制了放療在年輕有生育要求的宮頸癌患者的應(yīng)用。鑒于宮頸癌對(duì)化療僅中度敏感,所以,目前化療主要作為宮頸癌晚期患者的輔助性及姑息性治療手段。因此,尋找治療宮頸癌的新途徑及新方法具有顯著的意義。眾所周知,納米,作為一個(gè)長度計(jì)量單位,僅為十億分之一米。當(dāng)物質(zhì)進(jìn)入納米尺度,則會(huì)出現(xiàn)明顯的性能方面的改變。而納米醫(yī)學(xué),則是把納米科學(xué)技術(shù)與現(xiàn)代醫(yī)學(xué)的研究融合在一起新興學(xué)科。在納米治療的領(lǐng)域當(dāng)中,納米藥物表現(xiàn)出傳統(tǒng)化療藥物所不具備的優(yōu)點(diǎn),如粒徑小、滲透能力強(qiáng)等,抗腫瘤納米藥物既可直接作為抗腫瘤藥物作用于腫瘤組織,也可經(jīng)改造為載藥體系,在細(xì)胞水平和動(dòng)物水平均表現(xiàn)出良好的腫瘤治療應(yīng)用前景。目前大量的研究證實(shí),納米粒可通過干擾參與細(xì)胞增殖、凋亡、轉(zhuǎn)錄因子等相關(guān)蛋白的表達(dá),從而作用于特定的信號(hào)轉(zhuǎn)導(dǎo)通路,發(fā)揮出特異的生物學(xué)活性。金納米粒(Au NPs)可抑制MAPK信號(hào)通路從而抑制腫瘤的增殖;納米氧化鋅(Zn O)可通過抑制Akt、Erk1/2的磷酸化,激活JNK、P38等,抑制腫瘤的轉(zhuǎn)移。大量的研究表明納米藥物可作用于多條信號(hào)轉(zhuǎn)導(dǎo)通路,具有選擇性的藥理活性,因此,納米抗腫瘤藥物具有深入探索和研究的巨大潛力和價(jià)值。氧化亞銅納米粒(Cuprous Oxide Nanoparticles,CONPs)是一種含銅納米藥物,分子式為Cu2O,目前已有研究表明,氧化亞銅納米粒CONPs可抑制惡性腫瘤細(xì)胞的增殖和轉(zhuǎn)移,并且研究證實(shí)氧化亞銅納米粒CONPs可通過線粒體介導(dǎo)的細(xì)胞凋亡信號(hào)通路發(fā)揮抗腫瘤的作用。本課題通過cck8實(shí)驗(yàn),transwell實(shí)驗(yàn),細(xì)胞凋亡、細(xì)胞周期檢測,發(fā)現(xiàn)氧化亞銅納米粒CONPs可顯著抑制宮頸癌細(xì)胞的增殖,遷移和侵襲,并且可誘導(dǎo)宮頸癌細(xì)胞凋亡,阻滯宮頸癌細(xì)胞周期。通過裸鼠荷瘤實(shí)驗(yàn)發(fā)現(xiàn),氧化亞銅納米粒CONPs不僅可抑制裸鼠腫瘤的增殖,且與順鉑治療組相比,氧化亞銅納米粒CONPs治療組無明顯體重下降等副作用。體內(nèi)外結(jié)果實(shí)驗(yàn)都提示著氧化亞銅納米粒CONPs在宮頸癌治療中有著良好的應(yīng)用前景。為了更好的研究氧化亞銅納米粒CONPs治療宮頸癌的作用機(jī)制,為臨床轉(zhuǎn)化應(yīng)用打下堅(jiān)實(shí)的基礎(chǔ),我們通過透射電鏡(TEM)分析發(fā)現(xiàn),氧化亞銅納米粒CONPs可穿過細(xì)胞膜,聚集在細(xì)胞的線粒體,內(nèi)質(zhì)網(wǎng)等部位。并且發(fā)現(xiàn),隨著氧化亞銅納米粒CONPs濃度的增大,細(xì)胞自噬體的產(chǎn)生也隨之增多。通過激光共聚焦及流式分析發(fā)現(xiàn),氧化亞銅納米粒CONPs可損傷線粒體導(dǎo)致線粒體膜電位下降,并且通過自噬流研究也進(jìn)一步確認(rèn)了隨著氧化亞銅納米粒CONPs濃度的增大,細(xì)胞內(nèi)自噬體的產(chǎn)生是隨之增多的。通過Western Blot檢測LC3等自噬相關(guān)蛋白,結(jié)果也與之前相符,即隨著氧化亞銅納米粒CONPs處理濃度的增大,LC3 II/I的比例也隨之增加,并且,隨著氧化亞銅納米粒CONPs處理時(shí)間的延長,LC3 II/I的比例也有著一定程度的增加。鑒于目前自噬的發(fā)生與AKT/m TOR通路有著密切的關(guān)系,我們檢測了相關(guān)蛋白的表達(dá),實(shí)驗(yàn)結(jié)果分析發(fā)現(xiàn),CONPs可通過抑制AKT/m TOR通路,降低AKT、m TOR的磷酸化水平,從而發(fā)揮抗腫瘤的作用。綜上所述,本課題研究表明CONPs可抑制宮頸癌的增殖,誘導(dǎo)宮頸癌細(xì)胞的凋亡,并且可通過抑制AKT/m TOR通路發(fā)揮作用。
[Abstract]:Cervical cancer is one of the most common malignant tumors in women. Nearly 500 thousand of women in the world are diagnosed with cervical cancer in the world every year. In China, the incidence and mortality of the women are nearly 1/4 worldwide, which seriously threaten the health of women in our country. At present, the incidence of cervical cancer is closely related to the high-risk type of human papillomavirus (HPV, Human papillomavirus). Statistics show that HPV16,18,31,33,35,39,45,51,52,56,58,59,68,73 and type 82 are all related to cervical cancer, and the infection rate of type HPV16 and type 18 in cervical cancer is about 50% and 20% respectively. A large number of research data show that the E6 and E7 protein in the high risk type of HPV plays a vital role in the maintenance of cervical cancer, of which the high risk is at high risk. Type HPV E6 can induce the degradation of tumor suppressor factor p53 through ubiquitination, so as to eliminate its growth inhibition signal.E7, which can phosphorylate Rb protein, and thus play a role in promoting tumor proliferation. At present, the main treatment methods of cervical cancer are operation and radiotherapy and chemotherapy. The operation is mainly applied to early patients with cervical cancer. Although radiotherapy is applicable to all stages of cervical cancer, the treatment is suitable for all stages. Patients, but because radiotherapy can cause irreversible damage to the vagina and ovary, and therefore, it also restricts the use of radiotherapy in young patients with reproductive requirements for cervical cancer. In view of the moderate sensitivity of cervical cancer to chemotherapy, chemotherapy is currently used as an auxiliary and palliative treatment for advanced cervical cancer patients. New approaches and methods for cervical cancer are significant. As we all know, nanoscale, as a measurement unit of length, is only 1/1000000000 meters. When material enters nanoscale, there will be obvious changes in performance. Nanomedicine is a new subject that combines nanotechnology and modern medicine. In the field of nanotherapy, nano drugs show the advantages that traditional chemotherapeutic drugs do not possess, such as small size and strong osmosis. Antitumor nanodrugs can be used as antitumor drugs directly to tumor tissue, and can also be transformed into drug loading systems, and good tumor treatment applications are shown at both the cell level and the animal level. A large number of studies have confirmed that nanoparticles can interfere with the expression of cell proliferation, apoptosis, transcription factors and other related proteins by interfering with specific signal transduction pathways, and play a specific biological activity. Au NPs can inhibit MAPK signaling and inhibit the proliferation of tumor; nano Zinc Oxide (Zn O) can be used. Excessive inhibition of Akt, Erk1/2 phosphorylation, activation of JNK, P38, and so on, inhibit tumor metastasis. A large number of studies have shown that nanometers can play a role in multiple signal transduction pathways and have selective pharmacological activity. Therefore, nano antitumor drugs have great potential and value for exploration and research. Cuprous Oxide Nanoparticl (Cuprous Oxide Nanoparticl) Es, CONPs) is a copper containing nano drug and molecular formula is Cu2O. Currently, it has been shown that cuprous nanoparticles CONPs can inhibit the proliferation and metastasis of malignant tumor cells, and the study confirms that the cuprous nanoparticles CONPs can play an antitumor effect through mitochondrial mediated apoptosis signal. This subject is conducted through CCK8 experiment, tr Answell experiments, apoptosis and cell cycle detection showed that CONPs nanoparticles could significantly inhibit the proliferation, migration and invasion of cervical cancer cells, and induce cervical cancer cell apoptosis and block cervical cancer cell cycle. It was found that oxidized cuprous nanoparticles CONPs can not only inhibit the proliferation of nude mice, but also inhibit the proliferation of tumor in nude mice. Compared with cisplatin treatment group, there is no obvious side effect in the treatment group of cuprous oxide nanoparticles CONPs. Both in vivo and in vivo results suggest that cuprous nanoparticles CONPs has a good prospect in the treatment of cervical cancer. In order to better study the mechanism of the effect of cuprous nanoparticles CONPs in the treatment of cervical cancer, it can be used for clinical transformation. On a solid basis, we found through transmission electron microscopy (TEM) analysis that the cuprous nanoparticles CONPs can pass through the cell membrane and gather in the mitochondria, endoplasmic reticulum and other parts of the cell. And it is found that the production of the cell autophagic increases with the increase of the concentration of cuprous oxide nanoparticles CONPs. The cuprous nanoparticles CONPs can damage mitochondria and lead to the decrease of mitochondrial membrane potential and further confirm that the production of autophagic in the cell is increased with the increase of CONPs concentration of cuprous oxide nanoparticles. The detection of autophagic related proteins such as LC3 by Western Blot is also consistent with the results before. With the increase of CONPs concentration of cuprous nanoparticles, the proportion of LC3 II/I increased, and the proportion of LC3 II/I increased with the prolongation of the CONPs treatment time of cuprous nanoparticles. In view of the close relationship between the occurrence of autophagy and the AKT/m TOR pathway, we detected the expression of the related proteins. The experimental results show that CONPs can reduce the phosphorylation level of AKT, m TOR by inhibiting the AKT/m TOR pathway, and thus exerts anti-tumor effect. In summary, this study shows that CONPs can inhibit the proliferation of cervical cancer and induce apoptosis of cervical cancer cells, and can play a role in the inhibition of AKT/m TOR pathway.

【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R737.33

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本文編號(hào)：1807934