Yuan Ling, Li Jia-Xin, Yang Yi, Chen Yan, Ma Ting-Ting, Liang Shuang, Bu Yang, Yu Lei, Nan Yi
Pharmacy College of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China.
Traditional Chinese Medicine College, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China.
World J Gastroenterol. 2021 Apr 28;27(16):1785-1804. doi: 10.3748/wjg.v27.i16.1785.
Gastric carcinoma (GC) is a digestive system disease with high morbidity and mortality. However, early clinical detection is difficult, and the therapeutic effect for advanced disease is not satisfactory. Thus, finding new tumor markers and therapeutic targets conducive to the treatment of GC is imperative. MRPL35 is a member of the large subunit family of mitochondrial ribosomal protein. MRPL35 shows the characteristic of oncogene in colorectal cancer and esophageal cancer, which promotes the exploration of the correlation between MRPL35 and GC. We proposed that the expression of MRPL35 might be critical in GC.
To study the effect of knockdown on GC cell proliferation.
The expression of in GC was evaluated based on data from the public tumor database UALCAN (www.ualcan.path.uab.edu). The effect of the expression of on the prognosis was evaluated with KMplot (www.kmplot.com). The expression of MRPL35 was assessed on the tissue microarray by immunohistochemistry and the level of mRNA in 25 pairs of clinical GC tissues and matched adjacent tissues was detected by quantitative reverse transcription-polymerase chain reaction. Celigo cell count assay, colony formation assay, and flow cytometry were used to assess the role of MRPL35 in GC cell proliferation and apoptosis . Additionally, tumor formation experiment in BALB/c nude mice was utilized to determine the effect of on GC cell proliferation. After knockdown of , related proteins were identified by isobaric tags for relative and absolute quantification analysis, and the expression of related proteins was detected by Western blot.
The expression of MRPL35 was up-regulated in GC ( = 1.77 × 10). The Kaplan-Meier plots of the overall survival indicated that high expression of MRPL35 was associated with a poor survival in GC. Compared with adjacent tissues, the expression of MRPL35 in GC tissues was increased, which was related to age ( = 0.03), lymph node metastasis ( = 0.007), and pathological tumor-node-metastasis stage ( = 0.024). Knockdown of inhibited GC cell proliferation and colony formation and induced apoptosis. Animal experiment results showed that knockdown of inhibited tumor formation in BALB/c nude mice. Western blotting analysis showed that after knockdown of , the expression of PICK1 and BCL-XL proteins decreased, and that of AGR2 protein increased.
Collectively, our findings demonstrate that knockdown of inhibits GC cell proliferation through related proteins including PICK1, BCL-XL, and AGR2.
胃癌（GC）是一种发病率和死亡率都很高的消化系统疾病。然而，早期临床检测困难，晚期疾病的治疗效果也不尽人意。因此，寻找有利于胃癌治疗的新肿瘤标志物和治疗靶点势在必行。MRPL35是线粒体核糖体蛋白大亚基家族的成员。MRPL35在结直肠癌和食管癌中表现出癌基因的特征，这促进了对MRPL35与胃癌相关性的探索。我们推测MRPL35的表达在胃癌中可能至关重要。
研究敲低MRPL35对胃癌细胞增殖的影响。
基于公共肿瘤数据库UALCAN（www.ualcan.path.uab.edu）的数据评估MRPL35在胃癌中的表达。用KMplot（www.kmplot.com）评估MRPL35表达对预后的影响。通过免疫组织化学在组织芯片上评估MRPL35的表达，并通过定量逆转录-聚合酶链反应检测25对临床胃癌组织及匹配的癌旁组织中MRPL35 mRNA的水平。使用Celigo细胞计数法、集落形成试验和流式细胞术评估MRPL35在胃癌细胞增殖和凋亡中的作用。此外，利用BALB/c裸鼠体内成瘤实验确定敲低MRPL35对胃癌细胞增殖的影响。敲低MRPL35后，通过相对和绝对定量的等压标签鉴定相关蛋白，并通过蛋白质免疫印迹法检测相关蛋白的表达。
MRPL35在胃癌中的表达上调（=1.77×10）。总生存的Kaplan-Meier曲线表明，MRPL35高表达与胃癌患者较差的生存率相关。与癌旁组织相比，MRPL35在胃癌组织中的表达增加，这与年龄（=0.03）、淋巴结转移（=0.007）和病理肿瘤-淋巴结-转移分期（=0.024）有关。敲低MRPL35可抑制胃癌细胞增殖和集落形成，并诱导细胞凋亡。动物实验结果表明，敲低MRPL35可抑制BALB/c裸鼠体内肿瘤形成。蛋白质免疫印迹分析表明，敲低MRPL35后，PICK1和BCL-XL蛋白表达降低，AGR2蛋白表达增加。
总的来说，我们的研究结果表明，敲低MRPL35通过PICK1、BCL-XL和AGR2等相关蛋白抑制胃癌细胞增殖。
