Department of Pharmacy, Yantai Yuhuangding Hospital, Yantai, China.
Eur Rev Med Pharmacol Sci. 2018 May;22(10):3010-3017. doi: 10.26355/eurrev_201805_15058.
To observe the effect of paclitaxel on the autophagy of human cervical cancer cell lines by the expression regulation of lncRNARP11-381N20.2 as well as to explore the interaction and relationship between autophagy, proliferation, and apoptosis of cervical cancer cells.
Genome-wide expression profiles of tumors and their susceptibility to drugs were downloaded through TCGA database to find differentially expressed lncRNA RP11-381N20.2 in chemosensitive sensitive and insensitive groups. Expression of RP11-381N20.2 in 60 cervical cancer tissues and 30 normal tissues was detected by qRT-PCR. The relationship between the expression of RP11-381N20.2 and the clinicopathological parameters of lung cancer was statistically analyzed. The recombinant plasmid pcDNA-RP11-381N20.2 and pcDNA-NC of RP11-381N20.2 were transfected into SiHa cells by lipofectamine, respectively. The autophagy and phenotypic effects were observed. Cell proliferation was determined by colony formation assay. Apoptosis was detected by flow cytometry. Western blot was conducted to detect expressions of autophagy-related proteins.
Genome-wide expression profiles of chemotherapy-sensitive and insensitive data in patients with cervical cancer in TCGA database were analyzed by edger package, results showed that the expression of lncRNA RP11-381N20.2 was significantly lower in the chemotherapy-insensitive group. qRT-PCR results showed that the expression of RP11-381N20.2 in cervical cancer was decreased, and the total survival time of patients was positively correlated with the expression of RP11-381N20.2. RP11-381N20.2 was associated with TNM (tumor node metastasis) staging and tumor size. Biological functions of SiHa cells showed that the expression of RP11-381N20.2 was negatively correlated with the treatment time and dose of paclitaxel. Colony formation assay showed that paclitaxel could inhibit the proliferation of cervical cancer cells in a dose-dependent manner. Flow cytometry showed that paclitaxel induced apoptosis of cervical cancer cells, which was more promoted after combination with RP11-381N20.2. Western blot results suggested that paclitaxel could induce autophagy in cervical cancer cells in a time- and dose-dependent manners. Paclitaxel combined with RP11-381N20.2 could significantly increase apoptosis of cervical cancer cells.
During the killing process of paclitaxel on cervical cancer SiHa cells, cell autophagy would affect the efficacy, after overexpression of RP11-381N20.2 in SiHa cells, autophagy induced by paclitaxel was inhibited, thereby enhancing the killing effect of paclitaxel on tumor cells.
观察长链非编码 RNA（lncRNA）RP11-381N20.2 通过调节紫杉醇对人宫颈癌细胞自噬的影响，探讨宫颈癌细胞自噬、增殖和凋亡之间的相互作用和关系。
通过 TCGA 数据库下载肿瘤全基因组表达谱及其对药物的敏感性，以找到化疗敏感性和耐药性组之间差异表达的 lncRNA RP11-381N20.2。采用 qRT-PCR 检测 60 例宫颈癌组织和 30 例正常组织中 RP11-381N20.2 的表达。统计分析 RP11-381N20.2 表达与肺癌临床病理参数的关系。通过脂质体将 RP11-381N20.2 的重组质粒 pcDNA-RP11-381N20.2 和 pcDNA-NC 转染 SiHa 细胞，观察自噬和表型效应。通过集落形成实验测定细胞增殖。通过流式细胞术检测细胞凋亡。Western blot 检测自噬相关蛋白的表达。
通过 edger 包分析 TCGA 数据库中宫颈癌化疗敏感和耐药数据的全基因组表达谱，结果表明 lncRNA RP11-381N20.2 在化疗耐药组中的表达明显降低。qRT-PCR 结果显示，RP11-381N20.2 在宫颈癌中的表达降低，且患者的总生存时间与 RP11-381N20.2 的表达呈正相关。RP11-381N20.2 与 TNM（肿瘤淋巴结转移）分期和肿瘤大小有关。SiHa 细胞的生物学功能显示，RP11-381N20.2 的表达与紫杉醇的治疗时间和剂量呈负相关。集落形成实验表明，紫杉醇能够以剂量依赖的方式抑制宫颈癌细胞的增殖。流式细胞术显示，紫杉醇诱导宫颈癌细胞凋亡，与 RP11-381N20.2 联合使用后凋亡更明显。Western blot 结果表明，紫杉醇能够以时间和剂量依赖的方式诱导宫颈癌细胞自噬。紫杉醇联合 RP11-381N20.2 可显著增加宫颈癌细胞的凋亡。
在紫杉醇对宫颈癌 SiHa 细胞的杀伤过程中，细胞自噬会影响疗效，过表达 SiHa 细胞中的 RP11-381N20.2 后，紫杉醇诱导的自噬受到抑制，从而增强了紫杉醇对肿瘤细胞的杀伤作用。
