【作者】 刘莹；

【导师】 梅金红；

【作者基本信息】 南昌大学 ， 临床医学硕士（专业学位）， 2023， 硕士

【摘要】 目的:研究白花蛇舌草对肺腺癌A549、H1975细胞株增殖和侵袭的影响及其可能机制,并探讨白花蛇舌草与吉西他滨抑制肺腺癌增殖和侵袭的协同作用。方法:1.通过CCK-8法检测不同浓度白花蛇舌草对肺腺癌A549、H1975细胞株增殖的影响。2.通过划痕实验和Transwell实验检测白花蛇舌草对各组细胞迁移、侵袭的影响。3.通过流式细胞术检测白花蛇舌草对各组细胞凋亡的影响。4.通过RT-q PCR和Western blot法检测白花蛇舌草干预后各组细胞Nrf2、HO-1 m RNA及蛋白的表达。5.通过CCK-8法检测吉西他滨对肺腺癌A549、H1975细胞株增殖的影响。6.白花蛇舌草和吉西他滨联合作用于肺腺癌A549、H1975细胞株,使用CCK-8法、划痕实验和Transwell实验检测各组细胞的增殖、迁移和侵袭情况。结果:1.CCK-8法结果显示:白花蛇舌草干预组肺腺癌A549、H1975细胞株的增殖受到抑制,且与其剂量呈时间-浓度依赖作用,差异具有统计学意义(P<0.05)。2.划痕实验和Transwell实验表明,和对照组相比,白花蛇舌草可抑制各组细胞的迁移和侵袭,差异有统计学意义(P<0.05)。3.流式细胞术实验结果表明,和对照组相比,白花蛇舌草可促进A549、H1975细胞株凋亡,差异有统计学意义(P<0.05)。4.RT-q PCR和Western blot实验提示,经白花蛇舌草干预后,各组细胞的Nrf2、HO-1 m RNA和蛋白的表达均降低,差异有统计学意义(P<0.05)。5.CCK-8法结果显示:吉西他滨可抑制肺腺癌A549、H1975细胞株的增殖,且与其剂量呈时间-浓度依赖作用,差异有统计学意义(P<0.05)。6.联合应用白花蛇舌草和吉西他滨后,在A549组中,与单用白花蛇舌草或单用吉西他滨相比,联合用药组细胞抑制率在48h时分别提高27.0%(P<0.01)和17.7%(P<0.05);迁移率分别下降32.1%(P<0.001)和26.4%(P<0.05);侵袭率分别下降28.1%(P<0.001)和22.5%(P<0.001)。在H1975组中,与单用白花蛇舌草或单用吉西他滨相比,联合用药组细胞抑制率在48h时分别提高33.7%(P<0.001)和64.6%(P<0.001);迁移率分别下降18.0%(P<0.001)和22.7%(P<0.001);侵袭率分别下降32.0%(P<0.001)和38.1%(P<0.001)。结论:1.白花蛇舌草可抑制肺腺癌A549、H1975细胞的增殖和侵袭。2.白花蛇舌草对肺腺癌A549、H1975细胞的抑制作用可能是通过调控Nrf2/HO-1信号通路完成的。3.白花蛇舌草和吉西他滨对肺腺癌A549、H1975细胞的增殖和侵袭的抑制作用具有协同性。更多还原

【Abstract】 Objective:To study the effect of Hedyotis diffusa Willd on the proliferation and invasion of lung adenocarcinoma A549 and H1975 cell lines and its possible mechanism,and to study the synergistic effect of Hedyotis diffusa Willd and Gemcitabine in inhibiting the proliferation and invasion of Lung adenocarcinoma.Methods:1.The effects of different concentrations of Hedyotis diffusa Willd on the proliferation of lung adenocarcinoma A549 and H1975 cell lines were detected by CCK-8 assay.2.The effects of Hedyotis diffusa Willd on the migration and invasion of each group of cells were detected by Wound healing test and Transwell chamber assay.3.The effect of Hedyotis diffusa Willd on apoptosis of each group of cells was detected by flow cytometry.4.RT-q PCR and Western blot were used to detect the levels of m RNA and protein expression of Nrf2 and HO-1 in each group of cells after treating with Hedyotis diffusa Willd.5.The effect of Gemcitabine on the proliferation of lung adenocarcinoma A549 and H1975 cell lines was detected by CCK-8 assay.6.The effect of the combination of Hedyotis diffusa Willd and Gemcitabine on the proliferation and invasion of lung adenocarcinoma A549 and H1975 cell lines was detected by CCK-8 assay,Wound healing test and Transwell chamber assay.Results:1.The results of CCK-8 assay showed that the proliferation of lung adenocarcinoma A549 and H1975 cell lines in the intervention group of Hedyotis diffusa Willd was inhibited and showed a time-concentration-dependent effect with its dose,and the difference was statistically significant(P < 0.05).2.Wound healing assay and Transwell assay showed that compared with the control group,Hedyotis diffusa Willd could inhibit the migration and invasion of cells in each group,and the difference was statistically significant(P < 0.05).3.The results of flow cytometry showed that compared with the control group,Hedyotis diffusa Willd could promote apoptosis of A549 and H1975 cell lines,and the difference was statistically significant(P < 0.05).4.RT-q PCR and Western blot suggested that the expression levels of Nrf2,HO-1m RNA and protein were reduced in all groups of cells after the intervention of Hedyotis diffusa Willd,and the difference was statistically significant(P < 0.05).5.The results of cell proliferation assay showed that Gemcitabine could inhibit the proliferation of lung adenocarcinoma A549 and H1975 cell lines,and it showed a time-concentration-dependent effect with its dose,and the difference was statistically significant(P < 0.05).6.After combined application of Hedyotis diffusa Willd and Gemcitabine,in the A549 group,compared with Hedyotis diffusa Willd alone or Gemcitabine alone,the cell inhibition rate at 48 h was increased by 27.0%(P < 0.01)and 17.7%(P < 0.05)in the combined group;the migration rate was decreased by 32.1%(P < 0.001)and26.4%(P < 0.05);the invasion rate was decreased by 28.1%(P < 0.001)and 22.5%(P < 0.001).In the H1975 group,compared with Hedyotis diffusa Willd alone or Gemcitabine alone,the cell inhibition rate at 48 h was increased by 33.7%(P < 0.001)and 64.6%(P < 0.001)in the combination group;the migration rate was decreased by18.0%(P < 0.001)and 22.7%(P < 0.001);and the invasion rate was decreased by32.0%(P < 0.001)and 38.1%(P < 0.001).Conclusions:1.Hedyotis diffusa Willd may inhibit the proliferation and invasion of lung adenocarcinoma A549 and H1975 cells.2.The inhibitory effect of Hedyotis diffusa Willd on lung adenocarcinoma A549 and H1975 cells may be accomplished by regulating the Nrf2/HO-1 signaling pathway.3.The inhibitory effects of Hedyotis diffusa Willd and Gemcitabine on the proliferation and invasion of lung adenocarcinoma A549 and H1975 cells were synergistic.更多还原