| Rhizoma Wenyujin Concisum is one of the traditional Chinese medicine for treating chronic hepatitis, liver cirrhosis and liver cancer. A great quantity studies demonstrated that curcumin, the extract of Curcuma longa, Calamus, turmeric and zedoary, can inhibite cell proliferation and promoting apoptosis of tumor cells. Previous studies show that curcumin can inhibit the malignant biological behavior of hepatocellular carcinoma cells by up-regulation of lncRNA AK125910, so as to control the development of liver cancer.AimOn the basis of the regulation lncRNA by curcumin with lncRNA expression microarray, the regulation of lncRNA AK125910 in HCC and the mechanism of curcumin intervention are further studied. It will reveal the relationship between lncRNA AK125910 and liver cancer. The relationship between lncRNA AK125910 and the effect of curcumin on HCC is also be studied.MethodLncRNA microarray was used to screen the lncRNA associated with HCC treated with carcinoma, and PCR and Western blot were used to verify. siRNA-AK125910 transiently transfected HCC is constructed to down-regulated lncRNA AK125910. MTT assay for cell apoptosis, Brdu mark for cell proliferation, and transewell assay for cell invasion are used in the study. Bioinformatics technique is used for detecting the target gene of AK125910 lncRNA. The expression of proteins related to the proliferation and invasion in HCC was detected in siRNA-AK125910 transiently transfected HCC.The resultsThere are 7 lncRNA (fold change>5.0) changed in HCC treated with curcumin by using lncRNA expression chips.5 incRNAs were upregulated, including AK125910, uc009ycs, HMlincRNA717, chrX:149438637-149438034 and chr7:155933475-155996100; and 2 IncRNAs were downregulated, including chrl:17129210938-17129213215 and AP1879664; which AK125910 upregulated most significant (9.78-fold). The expression of lncRNA AK125910 significantly increased by RT-PCR, for 8.92 ±2.33 times the control group (p <0.001), which is consistent with the microarray results show lncRNA AK125910 plays a regulatory role in apoptosis HCC treated with curcumin.MTT, brdu mark and Transwell assay were used in detection the role of lncRNA AK125910 in HCC treated with curcumin, it is confirmed the lncRNA AK125910 involved curcumin induces. It was confirmed that lncRNA AK125910 was involved apoptosis, proliferation and invasion and metastasis effect in HCC.MUC2 and SPAG4 are predicted to be the downstream target of incRNA AK125910 by bioinformatics method. And the enhancement effect on MUC2 and the inhibition effect on SPAG4 of lncRNA AK125910 are verified by Relatime PCR and western blot. LncRNA AK125910 can inhibit the expression of proliferation and invasion protein in HCC by RT-PCR and Western blot. LncRNA AK125910 involved in Bcl-2/BAX, caspase-3 and caspase-8, MMP-2 and MMP -9 expression.Conclusion1. The expression of AK125910 lncRNA in HCC cells was significantly increased, and curcumin could down-regulate AK125910 lncRNA in HCC cells2. Down-regulation of IncRNA AK125910 can inhibit the apoptosis, and promote proliferation and invasion in HCC. Curcumin promote apoptosis and inhibit proliferation and invasion by up-regulated lncRNAAK125910 in HCC.3. IncRNA AK125910 enhanced the expression of MUC2 and inhibited the expression of SPAG4. Also Bcl-2, BAX, Caspase-3, Caspase-8, MMP-2 and MMP-8 were involved.4. Bcl-2, MMP-2 and MMP-9 are decreased and Bax, caspase-3 and caspase-8 are increased by curcumin in HCC. Down-regulation IncRNA AK125910 can cancel the effects of curcumin. Curcumin induced apoptosis and inhibited of invasion by IncRNA AK125910 in HCC. |
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