miR-223-3p regulates proliferation and apoptosis of hepatocellular carcinoma SMMC-7721 cells by targeting RAC1 / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effects of miR-223-3p on the proliferation and apoptosis of hepatocellular carcinoma (HCC) cells by regulating Ras-related C3 botulinum toxin substrate 1 (RAC1) and its possible mechanism. Methods: Thirty pairs of HCC and corresponding para-cancer tissues resected in Jilin Central Hospital from August 2016 to August 2018 were collected for this study; in addition, human HCC cell lines SMMC-7721, BEL-7402, HepG2 and human normal hepatocyte QSG-7701 were also collected. The expression level of miR-223-3p in HCC tissue and cell lines was detected by qPCR. miR-223-3p mimics, miR-223-3p inhibitor and siRAC1 were transfected into SMMC-7221 cells, respectively. CCK-8 assay, Colony formation assay and Annexin V-FITC/PI staining Flow cytometry were used to detect the proliferation, clone formation and apoptosis of SMMC-7721 cells, respectively. The relationship between miR-223-3p and RAC1 was confirmed by Dual luciferase reporter gene assay. The protein level of RAC1 in SMMC-7721 cells was detected by Western blotting. Results: The expression of miR-223-3p in HCC tissues was significantly lower than that in paracaner tissues (P<0.01), and had significant correlation with pathological characteristics, such as tumor size, TNM stage, EdmondsonSteiner grade (all P<0.05 or P<0.01). miR-223-3p expression in HCC cell lines was significantly lower than that in QSG-7701 cells with the lowest expression in SMMC-7721 cells. Dual luciferase reporter gene assay confirmed that RAC1 was a target gene of miR-223-3p, and miR-223-3p negatively regulated RAC1 expression. Over-expression of miR-223-3p significantly inhibited the proliferation and colony formation (P<0.05 or P<0.01) of SMMC-7721 cells and promoted cell apoptosis (P<0.01). Contrarily, knockdown of miR-223-3p reversed the inhibitory effect of miR-223-3p mimics on cells. Conclusion: miR-223-3p over-expression inhibits proliferation and colony formation and promotes apoptosis of HCC cells, the mechanism of which may be related with its targeted down-regulation of RAC1.

Ampelopsin inhibited apoptosis and migration of SMMC-7721 cells through MAPKs signaling pathway / 药学学报

This research is aimed to investigate the effect of ampelopsin on apoptosis and migration of human hepatoma SMMC-7721 cells and explore the molecular mechanism. SMMC-7721 cells were pretreated with different doses of ampelopsin and cells proliferation was detected by CCK8 kit. Cell morphology was observed under an inverted microscope. Nuclear morphology was detected by DAPI staining. Apoptotic rate was detected by Annexin V-FITC/PI flow cytometry. Migration and invasion were detected by Transwell and scratch healing test. Western blotting was used to detect cleavage of poly ADP-ribose polymerase (PARP), expression of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), E-cadherin, and N-cadherin, and phosphorylation of ERK, P38 and JNK in MAPKs pathway. Our results showed that ampelopsin significantly inhibited proliferation and induced apoptosis of SMMC-7721 cells, with half inhibition dose (IC50) for 24 h was 38.98 μg·mL-1. With 50 μg·mL-1 ampelopsin treatment, typical apoptotic morphological changes occurred, such as cell detachment, shrinkage and nuclear condensation. Apoptotic rate increased from 15% to 55.1%, with PARP cleavage significantly increased. In addition, treatment of ampelopsin reduced scratch healing of cells and transmembrane cells number. The expression levels of MMP-2 and MMP-9 were decreased. Further analysis of EMT-related proteins showed that after ampelopsin treatment, E-cadherin was up-regulated and N-cadherin was down-regulated. During ampelopsin treatment, ERK reached its peak of activation after 1 h, while the maximum activation time of JNK was 12 h. Meanwhile, P38 was activated within 4 h, with the highest point at 2 h. But after 4 h, ampelopsin inhibited phosphorylation of P38. These results indicated that ampelopsin induced apoptosis and reduced migration through activating MAPKs pathway and reversing EMT process in SMMC-7721 cells. This work provides a mechanistic basis for utilizing ampelopsin for anti-hepatocarcinoma treatment.

Over-expression of PRRX1 induces apoptosis of hepatocellular carcinoma SMMC7721 cells through a p53-mediated mitochondrial apoptosis pathway / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of over-expression of paired related homoeobox 1 (PRRX1) on apoptosis of hepatocellular carcinoma SMMC7721 cells, and to explore its detailed mechanism. Methods： ：Lentivirus-mediated PRRX1 over-expression vector (pGMLV-PRRX1) and empty vector (Vector) were respectively infected into SMMC7721 cells, and the mRNA and protein expression levels of PRRX1 in infected cells were detected by qPCR and WB. The effect of PRRX1 over-expression on the cell proliferation and apoptosis of SMMC7721 cells were assessed by CCK-8 assay and Annexin-V FITC/PI double staining flow cytometry assay, respectively. The change of mitochondrial membrane potential of SMMC7721 cells was detected by mitochondrial membrane potential assay kit (JC-10 staining assay). The enzymatic activities of caspase-8 and caspase-9 in infected cells were detected by using caspase activity assay kit (spectrophotometric method). The protein expression levels of p53, Bcl-2, Bax, Fas, Cleaved-caspase-3 and Cty C expressed in mitochondria and cytosol were evaluated by WB. Results： ：PRRX1 over-expressed SMMC7721 cell line was successfully constructed, and the protein and mRNA expression levels of PRRX1 were significantly increased in lentivirus infected cells (all P< 0.01). Compared with control group and vector group, over-expression of PRRX1 significantly inhibited cell proliferation, weakened mitochondrial membrane potential, but increased the rate of apoptosis, elevated the shear level of caspase-3, promoted the release of Cyt C protein from mitochondrial into cytosol and increased the enzymatic activity of caspase-9 (all P<0.05 or P<0.01). In addition, over-expression of PRRX1 also promoted the protein expressions of p53 and Bax but inhibited the protein expression of Bcl-2 (all P< 0.05 or P<0.01); however,it had no significant effect on the expression of Fas protein and the enzymatic activity of caspase-8 (all P＞ 0.05). Conclusion： ：Over-expression of PRRX1 induces apoptosis in hepatocellular carcinoma SMMC7721 cells, which may be related to the activation of p53-mediated mitochondrial apoptotic pathway

Psoralen inhibits malignant proliferation and induces apoptosis through triggering endoplasmic reticulum stress in human SMMC7721 hepatoma cells

BACKGROUND: Psoralen is a coumarin-like and coumarin-related benzofuran glycoside, which is a commonly used traditional Chinese medicine to treat patients with kidney and spleen-yang deficiency symptom. Psoralen has been reported to show estrogen-like activity, antioxidant activity, osteoblastic proliferation accelerating activity, antitumor effects and antibacterial activity. However, the antitumor mechanism of psoralen is not fully understood. This study aimed to investigate the therapeutic efficacy of psoralen in human hepatoma cell line SMMC7721 and the mechanism of antitumor effects. RESULTS: Psoralen inhibited proliferation of SMMC7721 in a dose- and time-dependent manner, and promoted apoptosis. Further, psoralen activated the ER stress signal pathway, including the expansion of endoplasmic reticulum, increasing the mRNA levels of GRP78, DDIT3, ATF4, XBP1, GADD34 and the protein levels of GDF15, GRP78, IRE1α, XBP-1s in a time-dependent manner. Psoralen induces cell cycle arrest at G1 phase by enhancing CyclinD1 and reducing CyclinE1 expression. Moreover, TUDC couldn't inhibit the psoralen-induced ER stress in SMMC7721 cells. CONCLUSIONS: Psoralen can inhibit the proliferation of SMMC7721 cells and induce ER stress response to induce cell apoptosis, suggesting that psoralen may represent a novel therapeutic option for the prevention and treatment hepatocellular carcinoma.

Effect of myricetin on migation and invasion ofhepatoma SMMC-7721 cells / 中国药理学通报

Aim To investigate the effects of myricetin on the migration and invasion of hepatoma SMMC-7721 cells.Methods SMMC-7721 cells were treated with different concentrations of myricetin.Migration and invasion of SMMC-7721 cells were examined by Wound healing and Transwell, and the expression of E-cadherin and N-cadherin was detected by RT-qPCR and Western blot.Results Myricetin may inhibit the viability of SMMC-7721 cells via different concentration(10~40 μmol·L-1);furthermore, myricetin might also inhibit the migration and invasion of hepatoma SMMC-7721 cells.Meanwhile with the increasing of myricetin concentration, both filopodia and lamellipodia formation was reducd, and 7721 cells displayed in more integrity.And data from RT-qPCR and Western blot demonstrated that myricetin may up-regulate the expression of E-cadherin, simultaneously, down-regulate N-cadherin in SMMC-7721 cells.Conclusion Myricetin may influence the cell migration and invasion through up-regulating the expression of E-cadherin, simultaneously, down-regulating N-cadherin and activate cytoskeleton remolding of SMMC-7721.

Exposure of human hepatoma cells to nitrite and ammonia promotes invasive activity through activation of ROS/ODC pathway / 药学学报

Recent studies have demonstrated that nitrite and ammonia levels are higher in the tumor environment, but their effects on cancer cells remains unclear. The present study was designed to determine the effects of nitrite and ammonia on tumor invasion and the role of reactive oxygen (ROS)/ornithine decarboxylase (ODC) pathway. SMMC-7721 cells were treated with sodium nitrite, ammonium chloride, sodium nitrite and ammonium chloride mixture for 24 h, the cell viability was analyzed using the MTT assay, cell invasion was analyzed with the transwell assay, the intracellular ROS levels were detected with a reactive oxygen species (ROS) test kits, the expression of intracellular ODC was examined with immunofluorescence and Western blot, the expression of matrix metallopeptidase-2(MMP-2) and MMP-9 were analyzed by Western blot. Compared with the control group, SMMC-7721 cells exhibited an increase in cell viability, invasion ability, ROS levels and ODC protein after exposure to 150 μmol·L-1 sodium nitrite and ammonium chloride mixture for 24 h. The invasive activity was reduced by ROS scavenger N-acetycysteine (NAC) in SMMC-7721 cells. The specific ODC inhibitor difluoromethylornithine (DFMO) increased ROS levels and weakened the ability of sodium nitrite and ammonium chloride mixture in the regulation of invasion of SMMC-7721 cells. These data demonstrated that sodium nitrite and ammonium chloride mixture promote invasion of SMMC-7721 cells by enhancing ROS/ODC pathway.

Anticancer effects of crude extract from Melia toosendan Sieb. et Zucc on hepatocellular carcinoma in vitro and in vivo / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the anti-cancer effects of crude extract from Melia toosendan Sieb. et Zucc and its possible molecular mechanisms in vitro and in vivo.</p><p><b>METHODS</b>Transonic alcohol-chloroform extraction method was used to extract toosendanin from the bark of Melia toosendan Sieb. et Zucc, and the content of toosendanin in the crude extract was measured by high performance liquid chromatography (HPLC). Anti-cancer effects of crude extract from Melia toosendan Sieb. et Zucc were investigated in in vivo and in vitro studies. In the in vitro experiment, human hepatocellular carcinoma cell lines SMMC-7721 and Hep3B were co-incubated with toosendanin crude extract of different concentrations, respectively. In the in vivo experiment, BALB/c mice were subcutaneously inoculated with mouse hepatocellular carcinoma H22 cells and treated with crude extract.</p><p><b>RESULTS</b>HPLC revealed the content of toosendanin was about 15%. Crude extract from Melia toosendan Sieb. et Zucc inhibited cancer cells growth in a dose- and time-dependent manner. The 50% inhibitory concentration (IC50, 72 h) was 0.6 mg/L for SMMC-7721 cells and 0.8 mg/L for Hep3B cells. Both high-dose [0.69 mg/(kg d)] and low-dose [0.138 mg/(kg d)] crude extract could markedly suppress cancer growth, and the inhibition rate was greater than 50%. Hematoxylin and eosin staining showed necrotic area in cancers and transmission electron microscopy displayed necrotic and apoptotic cancer cells with apoptotic bodies. Immunohistochemistry showed that the expression of Bax and Fas increased and the expression of Bcl-2 reduced.</p><p><b>CONCLUSIONS</b>Toosendanin extract has potent anti-cancer effects via suppressing proliferation and inducing apoptosis of cancer cells in vivo and in vitro. The mechanism of apoptosis involves in mitochondrial pathway and death receptor pathway.</p>

SilencingIGF-1R gene inhibits proliferation of human SMMC7721 cell and promotes its apoptosis through down-regulatingBMP2 expression / 中国癌症杂志

Background and purpose:Insulin-like growth factor-1 (IGF-1) is a peptide that participates in many biological processes by stimulating the downstream signaling pathways through their interaction with IGF-1 re-ceptor (IGF-1R) and insulin receptor (IR). Bone morphogenetic proteins (BMPs) are a group of functional proteins which participate in the biological processes of proliferation and migration in many kinds of cancers and have become a hot area of cancer research. The study aimed to investigate the effects of silencingIGF-1R gene on the expression level ofBMP2 gene, and the cell proliferation and apoptosis of SMMC7721 cells.Methods:The RNAi plasmid targetingIGF-1R gene was constructed and transfected into SMMC7721 cells. Then the inhibition effect on the expression level of IGF-1R and BMP2 gene was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The SMMC7721 growth curve and cell apoptosis were detected by MTT assay and flow cytometry after they were transfected with RNAi plasmid.Results:The RNAi plasmid targetingIGF-1R gene was constructed successfully. The inhibition efficiencies at mRNA expression levels were 68.9% and 80.7% (IGF-1R gene), 79.5% and 83.3% (BMP2 gene), respectively, after transfection with IGF-1R-siRNA-1 and IGF-1R-siRNA-2 plasmid (P<0.05). The inhibition efficiencies at protein levels were 46.1% and 62.1% (IGF-1R gene,P<0.05), 42.5% and 60.9% (BMP2 gene,P<0.05), respectively. The results of MTT growth curve showed that the proliferation rate in the transfected SMMC7721 cells was significantly slower than that in the control group (P<0.05). The proportion of apoptotic cells in transfected groups was significantly higher than that in the control group (P<0.05).Conclusion:SilencingIGF-1R gene can downregulate the expression ofBMP2 gene at different levels that results in inhibition of cell proliferation and promotion of apoptosis in SMMC7721 cells.

The reverse effect of Gli2 gene silencing on epithelial-mesenchymal transition in hepatocellular carcinoma cell line SMMC-7721 / 基础医学与临床

Objective To explore the effect and mechanism of Gli2 on EMT and invasion in the hepatocellular car-cinoma cell line SMMC-7721.Methods shRNA of Gli2 and Negative control (NC) shRNA were constructed and transfected into SMMC-7721 cells.shRNA-Gli2 group,shRNA-NC group and blank group were set up .Transwell chambers assay , adhesion experiments were used to detect the ability of invasion ,homogeneous and heterogeneous cells intercellular adhesion of each group .Meanwhile, the qRT-PCR, Western blot were used to examine Gli2, E-cadherin ,N-cadherin and vimentin mRNA and protein expression .Results In different hepatocellular carcinoma cell lines and be along with the increasing ability of the invasion in hepatocellular carcinoma cell lines , Gli2 expres-sion was higher .Compared with the shRNA-NC group and blank control group ,the interfered group extensive cells invasion ability was inhibited ( P nism may be related to the up-regulation of E-cadherin and the dow N-regulation of N-cadherin, vimentin.

Evaluation of the hot and cold characteristics of ten antibiotics by cytological method / 西安交通大学学报(医学版)

Objective To evaluate the hot and cold characteristics of 10 antibiotics.Methods MTT assay was used to investigate the effects of 10 antibiotics on the growth and proliferation of cultured SMMC7721 cells and MFC-7 cells in vitro .Morphological changes were observed under the inverted microscope.Results The 10 antibiotics,namely,ampicillin,cefixime,cefpodoxime proxetil,cefaclor,cefalexin,azithromycin,clarithromycin, roxithromycin,doxycycline and oxytetracycline showed cold or cool characteristics.Morphological observation showed that cells treated with these drugs presented decreased cell density and turned round.Conclusion The results of this study demonstrated that the cytological method can be used to evaluate the hot and cold characteristics of western drugs.This simple and reliable method will promote research on Chinese medicalization of western drugs.

Inhibition of ursolic acid on angiogenesis and xenografts in zebrsfish (danio rerio) / 中国药理学通报

Aim To investigate the anti-angiogenesis and anti-xenograftes of UA in zebrafish larvaes. Meth-ods 24 hour post-fertilization ( hpf ) TG zebrafish was treated with different concentration of UA for 24h, and the number of intersegmental vessels( IVS) were detec-ted under fluorescent microscope respectively;then the models of AB/TG zebrafish xenografts were established by be micro-injected with SMMC-7721 or HT-29 cell at 48hpf respectively, and after cocultured with UA for 48h, optical density (OD) of the SMMC-7721 cell and subintestinal veins ( SIVs) induced by HT-29 were e-valuated under confocal microscope. Results ISV as-say showed that UA could cause IVS missing or disap-perance, inhibition ratio reaching 20. 25% and 26. 65%. UA blocked the spread of SMMC-7721 cells in zebrafish and OD value,and inhibition ratios at three concentrations were 38. 01%, 54. 69%, 61. 88%, re-spectively; in another SIVs assay induced by xeno-grafts, UA at concentration 10 and 15mg·L-1 showed that SIVs were inhibited (P<0. 01) obviously. Con-clusion UA could inhibit the angiogenesis and the growth of SMMC-7721/HT-29 xenografts,and the anti-tumor mechanism may be related with VEGFR2 expres-sion.

Effect of daidzein on hepatocellular carcinoma SMMC7721 cell proliferation and tumor stem cell CD133 expression / 中国生化药物杂志

Objective To discuss the effect of daidzein on hepatocellular carcinoma SMMC7721 cell proliferation CD133 expression on tumor stem cell.Methods Hepatocellular carcinoma SMMC7721 cells were cultured,digested and passaged,and divided into six groups with different drug:control group with no daidzein,100 μg/mL daidzein group,200μg/mL daidzein group,300μg/mL daidzein group,400μg/mL daidzein group ,500μg/mL daidzein.The inhibition ratio,hexokinase,alkaline phosphatase and CD133 levels in SMMC7721 cell were detected and compared at 24 h,48 h,72 h among those groups. Results The inhibition ratio was increased by Daidzein dose increasing,and decreased apparently by times extending,especially in 400μg/mL and 500μg/mL daidzeingroups.Compared with control group,the hexokinase and alkaline phosphatase activity and CD133 expression were decreased apparently in groups treated with daidzein(P<0.01).The more the dose,the higher the drop(P<0.01).Conclusion Daidzein can inhibit hepatocellular carcinoma SMMC7721 cell proliferation,and inhibit CD133 expression on tumor stem cell.

Survivin as a factor of radioresistance to high-LET carbon ions in human hepatoma SMMC.7721 cells / 中华放射医学与防护杂志

Objective To investigate the influences of survivin down-regulation on cell G2/M phase arrest,apeptosis and sensitivity to carbon ion irradiation. Methods Small interfering RNA (siRNA) targeting survivin mRNA was designed, in vitro chemo-synthesized and transfected into SMMC-7721 cells. Survivin mRNA expression in SMMC-7721 cells was measured by real-time PCR, and the apeptotic rates by Annexin-FTTC at 24 and 48 h after transfection. Cell G2/M phase arrest after transfection was assessed with flow eytometry as well. Cellular sensitivity to high-LET carbon ions was determined by means of colony-forming assay. Results The expressions of survivin at mRNA level were down-regulated to be 59% and 39% in relation to the non-treated cells at 24 and 48 h after siRNA transfeetion, respectively. G2/M phase arrest in SMMC-7721 cells at 24 h after transfection was observed while much more obvious at 48 h. The apeptotic rate of SMMC-7721 cells was 21.41 % at48 h after survivin siRNA transfection, which was significantly higher than that of the cells transfected with negative siRNA. Moreover, a decreased clonogenic survival in siRNA treated group was shown. Conclusion Down-regulation of survivin gene expression in SMMC-7721 cells by siRNA could effectively induce cell apeptosis and G2/M phase arrest, and enhance the cellular radiosensitivity to high-LET heavy ions.

Hairpin ribozyme targeted vascular endothelial growth factor (VEGF) inhibited the expression of VEGF and the growth of xenografted tumors / 肿瘤

Objective: Hepatocarcinoma cell line was transfected with anti-vascular endothelial growth factor (VEGF) hairpin ribozyme gene to observe the effect of hairpin ribozyme on VEGF expression and the growth of the xenografted tumors. Methods: The artificial anti-VEGF hairpin ribozyme gene was transfected into hepatocarcinoma SMMC-7721 cells via lipofectin mediation. The blank vector and the cell controls were prepared simultaneously. Then, positive clones were screened by genticin (G418). The transcription of ribozyme was confirmed by RT-PCR. The effects of the ribozyme on VEGF expression of SMMC-7721 cells were detected by semi-quantitative RT-PCR and immunohistochemical method. Cells in each group were inoculated into nude mice. The tumor volume and weight were recorded. The change in microvessel density and expression of VEGF was determined by immunohistochemistry. Results: Ribozyme gene was successfully transferred into tumor cells. The proliferation rate of ribozyme-transfected SMMC-7721 cells was significantly slower (P < 0.01). The expression of VEGF significantly decreased in ribozyme-transfected SMMC-7721 cells. After rebozyme transfection, the tumor formation rate significantly decreased and the growth speed of xenografted hepatocarcinoma markedly slowed down. The microvessel density and angiogenesis of the xenografted hepatocarcinoma were obviously reduced. Conclusion: Anti-VEGF hairpin ribozyme gene significantly inhibited the VEGF expression of hepatocarcinoma in vitro and in vivo by inhibiting angiogenesis of tumor cells. This study provided an experimental evidence for anti-angiogenesis gene therapy for hepatocarcinoma.

Anti-tumor Activity of Adriamycin-loaded Chitosan Nanoparticles Surface-modified with Glycyrrhizin on Human Hepatoma Cell Line SMMC-7721 / 中国药房

OBJECTIVE: To investigate the anti-tumor activity of adriamycin-loaded chitosan nanoparticles surface-modified with glycyrrhizin on human hepatoma cell line SMMC-7721. METHODS: Inhibition effect of drug-loaded nanoparticle on the proliferation of SMMC-7721 cells was determined with MTT method. Intra-cellular distribution of nanoparticles and morphological change of SMMC-7721 cells before and after treatment were observed by confocal laser scanning microscope (CLSM) and the apoptosis induced by adriamycin was measured by TUNEL assay. RESULTS: The inhibition effect of adriamycin-loaded chitosan nanoparticles surface-modified with glycyrrhizin on the proliferation of SMMC-7721 cells increased 6.36 times lower in comparison with free drugs. The inhibition effect of adriamycin-loaded chitosan nanoparticles had no obvious improvement. Nanoparticles surface-modified with glycyrrhizin promoted the distrubition of adriamycin in the nucleus, so as to promote the anti-tumor activity of adriamycin. TUNEL assay indicated that nanoparticles surface-modified with glycyrrhizin induce DNA fragmentation and nucleus breakdown to induce the apoptosis of SMMC-7721 cells. CONCLUSION: Adriamycin-loaded chitosan nanoparticles surface-modified with glycyrrhizin can used as potential active hepatocyte-targeted delivery carrier. Pharmacokinetic evaluation of it is worthy of further studing.

Periplocin of cortex periplocae inhibits Stat3 signaling and induces apoptosis in human hepatocellular carcinoma cell line SMMC-7721 / 第三军医大学学报

Objective To investigate the effect of periplocin of cortex periplocae (CPP) on Stat3 signaling and its probable molecular mechanism of inducing apoptosis and anti-tumor activity. Methods Cell proliferation was detected by MTT. Cell apoptosis and cell cycle were investigated by flow cytometry. Expression of Stat3 protein in SMMC-7721 cells was analyzed by Western blot. Mcl-1, Survivin and XIAP mRNA expressions were measured by RT-PCR. Results CPP inhibited the proliferation of SMMC-7721 cells significantly, induced their apoptosis and arrested their cell cycle at G2/M phase. Decreased expression of Stat3 protein in the cell nucleus was observed after CPP treatment, but no significant changes were found in cytoplasma. Mcl-1, Survivin and XIAP mRNA expression levels were decreased in a time-dependent manner. Conclusion CPP inhibits cell proliferation and induces apoptosis by inhibiting Stat3 signal transduction in human hepatocellular carcinoma cell line SMMC-7721.

siRNA targeting STAT5 induces apoptosis in human hepatocellular carcinoma cell line SMMC-7721 / 第三军医大学学报

Objective To investigate the effects of inhibition of STAT5 gene expression by RNA interference technology on apoptosis of human hepatocellular carcinoma cell line SMMC-7721. Methods Three siRNA eukaryotic expression vectors against STAT5 were constructed and transfected with lipofectamineTM 2000 into SMMC-7721 cells. The changes in STAT5 expression were detected by semi-quantitative RT-PCR and Western blot. Cell apoptosis was assayed by flow cytometry (FCM). Results The sequence-specific siRNA could effectively and specifically inhibit STAT5 gene expression at both mRNA and protein levels. The inhibition rates of STAT5 mRNA expression were 70.43%, 43.02%, and 45.07%, respectively. The inhibition rates of STAT5 protein expression were 67.45%, 37.36%, and 41.86%, respectively. At 48 h after transfection, apoptosis rate was 25.61%. Conclusion siRNA against STAT5 can inhibit STAT5 gene expression in SMMC-7721 cells effectively and specifically and induce apoptosis of SMMC-7721 cells. siRNA targeting STAT5 has a great potential value in gene therapy of hepatocellular carcinoma.