Yang Chao, Xue Jian-Guo
Department of Andrology, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210023 Jiangsu， China.
Department of Andrology, The Central Hospital of Enshi Tujia and Miao Autonomous Prefecture Enshi, 445000 Hubei, China.
Zhonghua Nan Ke Xue. 2023 Oct;29(10):881-887.
Exploring the effects and mechanisms of long non coding RNA (lncRNA) RPL22P1-201 on prostate cancer cell proliferation, cell cycle, and docetaxel sensitivity by regulating miR-216b-5p expression.
The Cancer LncRNA Census database was used to analyze the differential expression of RPL22P1-201 between prostate cancer tissue and normal tissue. Real time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression level of RPL22P1-201 in prostate cancer cell lines (DU-145, C4-2B, PC3, 22Rv1, LNCaP) and normal prostate epithelial cells (RWPE-1). PC3 cells were divided into si-RPL22P1-201 group (transfected with RPL22P1-201 interference sequence) and si-NC group (transfected with si-NC sequence). Colony formation assay was used to detect the proliferation ability of PC3 cells. Flow cytometry was used to detect the PC3 cell cycle. The CCK-8 method was used to detect the proliferation of PC3 cells in each group after treatment with docetaxel. The dual luciferase reporter gene experiment verifies the binding of RPL22P1-201 to the target gene. qRT-PCR was used to detect the expression level of miR-216b-5p. Western blot was used to detect the expression levels of TrkB, CDK4, cyclin D2, cyclin D3, and CDK6 proteins.
The expression level of RPL22P1-201 in prostate cancer tissue was higher than that in normal tissue (P<0.01). The expression level of RPL22P1-201 in prostate cancer cell lines was higher than that in normal prostate epithelial cells (P<0.01). The number of colonies in the si-NC group and si-RPL22P1-201 group was (256.1 ± 28.79) and (78.77 ± 14.52), respectively. The difference was statistically significant (P<0.01). The G0/G1 cell rates in the si-NC group and si-RPL22P1-201 group were (43.18 ± 4.56)% and (68.85 ± 3.40)%, respectively. The S cell rates were (36.84 ± 2.28)% and (24.27 ± 2.74)%, respectively. The G2/M cell rates were (19.98 ± 2.69)% and (6.88 ± 1.57)%, respectively, and the differences were statistically significant (all P<0.05). The cell survival rate of the si-RPL22P1-201 group under the action of docetaxel was lower than that of the si-NC group (all P<0.05). RPL22P1-201 can pair and bind with miR-216b-5p (P<0.01). Compared with the si-NC group, the si-RPL22P1-201 group showed a decrease in miR-216b-5p expression in PC3 cells (P<0.01), and a decrease in TrkB, CDK4, cyclin D2, cyclin D3, and CDK6 protein expression.
RPL22P1-201 is highly expressed in prostate cancer, and silencing RPL22P1-201 inhibits prostate cancer PC3 cell proliferation and cell cycle by increasing miR-216b-5p expression, and enhances PC3 cell sensitivity to docetaxel.
通过调控miR-216b-5p表达，探讨长链非编码RNA（lncRNA）RPL22P1-201对前列腺癌细胞增殖、细胞周期及多西他赛敏感性的影响及其机制。
利用癌症lncRNA普查数据库分析RPL22P1-201在前列腺癌组织与正常组织中的差异表达。采用实时定量聚合酶链反应（qRT-PCR）检测RPL22P1-201在前列腺癌细胞系（DU-145、C4-2B、PC3、22Rv1、LNCaP）及正常前列腺上皮细胞（RWPE-1）中的表达水平。将PC3细胞分为si-RPL22P1-201组（转染RPL22P1-201干扰序列）和si-NC组（转染si-NC序列）。采用集落形成试验检测PC3细胞的增殖能力。运用流式细胞术检测PC3细胞周期。采用CCK-8法检测多西他赛处理后各组PC3细胞的增殖情况。通过双荧光素酶报告基因实验验证RPL22P1-201与靶基因的结合。采用qRT-PCR检测miR-216b-5p的表达水平。运用蛋白质免疫印迹法检测TrkB、CDK4、细胞周期蛋白D2、细胞周期蛋白D3和CDK6蛋白的表达水平。
RPL22P1-201在前列腺癌组织中的表达水平高于正常组织（P<0.01）。RPL22P1-201在前列腺癌细胞系中的表达水平高于正常前列腺上皮细胞（P<0.01）。si-NC组和si-RPL22P1-201组的集落数分别为（256.1±28.79）和（78.77±14.52），差异具有统计学意义（P<0.01）。si-NC组和si-RPL22P1-201组的G0/G1期细胞率分别为（43.18±4.56）%和（68.85±3.40）%，S期细胞率分别为（36.84±2.28）%和（24.27±2.74）%，G2/M期细胞率分别为（19.98±2.69）%和（6.88±1.57）%，差异均具有统计学意义（均P<0.05）。si-RPL22P1-201组在多西他赛作用下的细胞存活率低于si-NC组（均P<0.05）。RPL22P1-201可与miR-216b-5p配对结合（P<0.01）。与si-NC组相比，si-RPL22P1-201组PC3细胞中miR-216b-5p表达降低（P<0.01），TrkB、CDK4、细胞周期蛋白D2、细胞周期蛋白D3和CDK6蛋白表达也降低。
RPL22P1-201在前列腺癌中高表达，沉默RPL22P1-201可通过增加miR-216b-5p表达抑制前列腺癌PC3细胞增殖和细胞周期，并增强PC3细胞对多西他赛的敏感性。
