探讨姜黄素抑制肝细胞癌（肝癌）索拉非尼耐药作用及其调控机制。

采用梯度浓度加药法成功获取人索拉非尼耐药肝癌细胞株Hep3B-SR，采用姜黄素20 μg/ml干预Hep3B-SR；长链非编码RNA（LncRNA）表达谱芯片筛选姜黄素干预Hep3B-SR的相关LncRNA；qRT-PCR检测靶标LncRNA和下游相关基因CD44、MYC、JUN、FOS mRNA变化情况；构建敲低和过表达靶标LncRNA细胞株，采用细胞克隆形成实验和细胞成球实验，检测肝癌干性变化情况；CCK-8检测索拉非尼IC₅₀值的变化情况；Western blot检测肝癌细胞干性标志物CD44及Wnt2、β-catenin、c-Myc蛋白及通路变化情况。两组细胞mRNA和蛋白表达比较采用t检验，多组肝癌细胞增殖和IC₅₀值比较采用单因素方差分析和LSD-t检验。

CCK8法检测显示，索拉非尼耐药株Hep3B-SR的平均IC₅₀值为（8.4±1.1）μmol，明显高于Hep3B细胞的（4.0±1.1）μmol（LSD-t=16.27，P<0.001）和姜黄素（20 μg/ml）干预后Hep3B-SR+Curcumin细胞的（6.1±1.1）μmol（LSD-t=3.97，P<0.001）。LncRNA表达谱芯片及qRT-PCR检测显示姜黄素可有效上调耐药肝癌细胞中LncCCDC152的表达，通过过表达LncCCDC152后可显著下调Hep3B肝癌细胞的成球和克隆形成能力；LncCCDC152可结合转录延伸因子1（TCERG1），进而影响肝癌细胞中Wnt/β-catenin通路基因的转录活性和蛋白表达。

姜黄素可抑制肝癌索拉非尼耐药，可能通过上调肝癌细胞中LncCCDC152结合TCERG1蛋白靶向抑制Wnt/β-catenin通路，从而减少肝癌干性活性并抑制索拉非尼耐药。

To evaluate the inhibitory effect of curcumin on sorafenib resistance in hepatocellular carcinoma (HCC) and unravel its regulatory mechanism.

Human sorafenib-resistant Hep3B-SR cell lines were successfully obtained by gradient concentration dosing method, and intervened by curcumin at a dose of 20 μg/ml. Long-chain non-coding RNA (LncRNA) expression microarray was used to screen the LncRNAs related to curcumin-intervened Hep3B-SR. qRT-PCR was employed to detect the changes of target LncRNA and the downstream related genes of CD44, MYC, JUN and FOS mRNA. The cell lines with knockdown and overexpression of target LncRNA were constructed. Cell colony and sphere formation assays were utilized to detect the stem-like changes of HCC. CCK-8 assay was adopted to detect the changes of IC₅₀ value of sorafenib. Western blot was performed to detect the changes of CD44, Wnt2, β-catenin, c-Myc proteins and the signaling pathway. The expression levels of mRNAs and proteins between two groups were compared by t test. The proliferation rate and IC₅₀ value of HCC cells among multiple groups were compared by one-way ANOVA and LSD-t test.

CCK8 assay showed that the average IC₅₀ value of sorafenib-resistant Hep3B-SR cell line was (8.4±1.1) μmol, significantly higher than (4.0±1.1) μmol of Hep3B cells (LSD-t=16.27, P<0.001) and (6.1±1.1) μmol of Hep3B-SR cells after curcumin (20 μg/ml) intervention (LSD-t=3.97, P<0.001). LncRNA expression microarray and qRT-PCR showed that curcumin could effectively up-regulate the expression of LncCCDC152 in drug-resistant HCC cells. Overexpression of LncCCDC152 could significantly down-regulate the sphere and colony formation ability of Hep3B HCC cells. LncCCDC152 could bind with transcriptional elongation regulator 1 (TCERG1), thus affecting the transcriptional activity and protein expression of the genes in the Wnt/β-catenin signaling pathway in cells.

Curcumin can suppress the sorafenib resistance of HCC, which targetedly inhibits the Wnt/β-catenin pathway probably by up-regulating the binding of LncCCDC152 with TCERG1 protein in HCC cells, thereby reducing stem-like activity of HCC and inhibiting sorafenib resistance.