Experimental study on effect of recombinant human growth hormone combined with chemotherapy on stomach neoplasms implanted in nude mice

Objective: To investigate the effect of different doses of recombined growth hormone (rhGH) on stomach neo- plasms implanted in nude mice, and its efficacy in combining with chemotherapy (flurouracil, 5-FU). Methods: Human stom- ach neoplasms model was established in nude mice. The nude mice were divided into control group, moderate-dose of rhGH group, low-dose rhGH group, 5-FU group, moderate-dose rhGH/5-FU group, and low-dose rhGH/5-FU group. The results of each group were observed after ten days. Results: After therapy, the body mass of rhGH groups was significantly increased compared with control group (P<0.05), the body mass of rhGH/5-FU groups was significantly increased compared with 5-FU group (P<0.05), but it was no significant difference between rhGH/5-FU groups and control group (P>0.05). The average tumor mass and volume of rhGH groups were not significantly increased compared with control group (P>0.05), but they were significantly reduced in 5-FU group and rhGH/5-FU groups (P<0.05). They were no significant difference between rhGH/5- FU groups and 5-FU group (P>0.05). After treatment, the percentages of S, G0/G1 and G2/M phases and proliferation index (PI) were not significantly changed in rhGH groups compared with control group (P>0.05), and the same with rhGH/5-FU groups compared with 5-FU group (P>0.05). The difference caused by dose of rhGH was not significant. Conclusion: rhGH enhances body mass, does not stimulate tumor growth, and has no adverse effects on tumor bearing nude mice. Combined with flurouracil, rhGH does not influence the efficacy of chemotherapy, and has no effect on tumor cell cycle kinetics.

Clinical observation of 125 I labeled anti alpha fetoprotein antibody radioimmunotherapy in hepatocellular carcinoma *

AIM To observe the therapeutic effects and toxic side reactions of 125 I labeled hourse anti human AFP polyclonal antibodies in immuno targeting therapy against hepatocellular carcinoma (HCC).

Prevention of metastasis to liver by using 5-FU intraperitoneal chemotherapy in nude mice inoculated with human colonic cancer cells

AIMS Using a new approach of regional adjuvant chemotherapy to prevent cancer cells hepatic metasta- sis after radical surgery of large bowel cancer. METHODS A model of liver with metastasis of hu- man colonic cancer (HCC) cells in nude mice was used to observe the effect in prevention of metastasis of HCC cells inoculated via spleen applied with early postoper- ative intraperitoneal (IP) chemotherapy using large dose of 5-FU. RESULTS The incidence of metastasis to liver was decreased by 40%,the mean number of metastatic liv- er nodules in each animal was reduced by 50.89% and the mean survival times of each animal was prolonged by 48.21% by using 5-FU 40 mg/NS 40 ml/kg IP for two consecutive days as compared with the controls. CONCLUSIONS IP is a new and more effective re- gional adjuvant chemotheraputic approach in the pre- vention of liver metastasis HCC cells after radical surgery of large bowel cancer.

Double-bullet radioimmunotargeting therapy in 31 patients with primary liver cancer

AIM To observe the effect of a double bullet immunotargeting therapy with the merit of chemotherapy and internal radiotherapy for primary liver cancer. METHODS The polyclonal horse antibody against human AFP (anti AFPAb) and the monoclonal murine antibody against human AFP (anti AFPMcAb) were used as carriers, and 131 I and mitomycin C (MMC) as warheads, to form double bullet, ie, 131 I anti AFPMcAb MMC (double bullet 1) and 131 I anti AFPAb MMC (double bullet 2) prepared by the modified chloramine T method. The double bullet targeting therapy was administered by intravenous drip once a month in 31 patients (treatment group) with unresectable primary liver cancer. Among them 4, 17 and 10 patients were administered 1, 2 and 3 times, and the median value of radiation dose (MBq/case) were 193 5±37 74; 651 9±232 4, and 992 0±230 5 respectively. RESULTS The shrinkage of tumor, AFP decrease and 1 and 2 year survival rates were significantly higher than those of the control groups of transarterial infusion (TAI) or transarterial chemoembolization (TACE) at the same time (50 0%, 15/30 vs 30 0%, 9/30, P <0 05; 66 7%, 18/27 vs 28 0%, 7/25, P <0 01 and 50 0%, 34 0% vs 33 0%, 3 3%, P <0 01, respectively). Furthermore, the tumor progression rate (10%) in treatment group was significantly lower than that of control group (40 0%, P <0 01). CONCLUSION Double bullet target therapy has a better effect due to the synergistic effects of antibody, radioisotope, and anticancer agents, thus enhancing the tumor killing effect.

Hepatic arterial infusion chemotherapy and embolization in the treatment of primary hepatic carcinoma

AIM To study the effects of hepatic arterial infusion chemotherapy (HAI) or embolization (HAE) in the treatment of primary hepatic carcinoma (PHC) and the factors influencing the effects. METHODS Follow up information in 188 patients (166 males and 22 females) with PHC treated with HAI ( n =82) or HAE ( n =106) were analyzed retrospectively. RESULTS The overall therapeutic effects were as follows: symptomatic relief in 59 6%; tumor shrinkage in 55%, blood AFP decrease in 37 8% and the overall survival rates of 1/2, 1, 2 and 3 years after treatment being 75 4%, 46%, 23 5% and 14 7%, respectively. The mean survival time was 12 2 months and the longest survival period was 50 months after treatment. In the HAI group the 1/2, 1, 2 and 3 years survival rates were 61 0%, 25 4%, 4 5% and 0%, respectively, the mean survival time being 7 7 months; in the HAE group the survival rates were 86 8%, 61 7%, 37 8% and 26 1%, respectively, the mean survival time being 15 7 months. Eighteen patients received secondary surgery. Seven factors influencing the therapeutic effects were analyzed. CONCLUSION Those cases of PHC with or without mild liver cirrhosis, stages Ⅰ and Ⅱ, single tumor, tumor size less than 10cm in diameter, no cancerous thrombus within the portal vein or hepatic arterio venous fistula, and HAE treatment time more than 3 times had better therapeutic effects than the contrast cases. To overcome these influencing factors and to adopt the comprehensive therapy would improve the effects of HAI or HAE in the treatment of PHC.

Comparative study on the effects of hepatic arterial embolization with Bletilla Striata or gelfom in the treatment of primary hepatic carcinoma

AIMS To study the effect and safety of hepatic arte- rial embolization (HAE) with Bletilla Striata powders of traditional Chinese drug in the treatment of primary hep- atic carcinoma (PHC). METHODS From May 1990 to September 1993,106 patients with PHC were treated by HAE with Bletilla Striata powders (n=56) or gelfoam powders (n=50) respectively under the controlled technical conditions. Authors analyzed the effects and complications in two groups. RESULTS Bletilla Striata powders produced exten- sive permanent proximal embolization of hepatic artery and collateral circulation formed. Intermission of treat- ment was as long as 6-12 months and tumor necrosis and shrinking were obvious. The survival rates of 1,2 and 3 years were 81.9%,44.9% and 33.6% respec- tively and the mean survival time was 19.8 months without serious complications. All clinical effects of Bletilla Striata group were better than those of glfoam group. CONCLUSIONS Bletilla Striata powders are superi- or to gelfoam powders as an angioembolus for hepatic carcinoma.

Protocol of Interventional Treatment for Hepatocellular Carcinoma

To establish a reasonable protocol for interventional treatment ofhcpatocellular carcinoma (HCC). Methods: The data of 1000 HCC patients treated by different kinds ofinterventional treatments were reviewed with their results of biochemistry, imaging, pathology andsurvival rate evaluated. The values as well as the pros and cons of these various kinds ofinterventional treatments were compared in order to find an optimal protocol. Results:Segmental-transcatheter oil chemoembolization (S-TOCE) could more effectively eradicate the tumoryet inflicting less damage on the noncancerous hepatic tissue and giving much higher survival ratethan the conventional transcatheter oil chemoembolization (C-TOCE). Precutaneous ethanol injection(PEI) in combination with chemoembolization could eliminate the residual tumor and significantlyincrease the survival rate without damaging the noncancerous hepatic tissue. The living quality orsurvival rate could be improved by choosing different ways of interventional treatments to cut downthe complications. Conclusion: The selection of different interventional treatments should bo doneaccording to the size and type of HCC. Active management is indicated for different complicationspresenting along with HCC.

Histopathological study of hepatocellular carcinoma after transcatheter hepatic arterial embolization

AIMS To study the histopathological changes in hepatocellular carcinoma (HCC) after transcatheter arterial embolization (TAE). METHODS Histopathological analysis was made in 39 cases of liver neoplasms after TAE and 11 cases of liver neoplasms after digital selective angiography (DSA), including pathological type, histological grade, necrotic degree, capsule, times of treatment, injured vessel and lymphocyte infiltration. RESULTS Six cases with 100% necrosis, 14 cases with 30% 95% necrosis, 19 cases with 0% 5% necrosis after TAE and 11 cases without necrosis after DSA were found histologically. The necrosis was related to the pathological type, capsule, injured vessels, but not to the histological grade, time of treatment and lymphocyte infiltration of the liver neoplasms. CONCLUSIONS TAE is an effective therapy for the late stage HCC. The encapsulated HCC is a preferable indicator for TAE.

Colon cancer and the immune system:The role of tumor invading T cells

Colon cancer is still one of the leading causes of cancer death worldwide. Although the host immune system has been shown to react against tumor cells, mainly through tumor infi ltrating lymphocytes and NK cells, tumor cells may utilize different ways to escape anti-tumor immune response. Tumor infi ltration of CD8+ and CD4+ (T-bet+) effector T cells has been attributed to a beneficial outcome, and the enhancement of T cell activation through T cell receptor stimulation and co-stimulatory signals provides promising strategies for immunotherapy of colon cancer. Growing evidence supports a role for the Fas/FasL system in tumor immunology, although the mechanisms and consequences of FasL activation in colon cancer are not completely understood. In animal models, depletion of regulatory T cells (CD4+ CD25+ T cells) can enhance the anti-tumor immune response under certain conditions. Taken together, recent insights in the immune reaction against colon carcinoma have provided new approaches to immunotherapy, although much remains to be learned about the exact mechanisms.

Clinical and experimental study on regional administration of phosphorus32 glassmicrospheres in treating hepatic carcinoma

AIM To study the therapeutical effectiveness, dosage range and toxic adverse effects of domestic phosphorus 32 glass microsphere and evaluate its clinical significance. METHODS Ⅰ. Fifty two BALB/*!c tumor bearing male nude mice were allocated into treatment group( n =38) and control group( n =14). In the former group different doses of 32 P GMS were injected into the tumor mass, while in the latter 31 P GMS or no treatment was given. The experimental animals were sacrificed in batches, and then the tumors and their nearby tissues were examined by light and electron microscopy. Ⅱ. Through selective catheterization of hepatic artery, 32 P GMS was infused to 5 healthy domestic pigs in a dosage equivalent to the therapeutic dose for human being, and 31 P GMS was infused to another 5 healthy domestic pigs. Two pigs infused with contrast medium served as whole course blank controls. One pig from each group was surrendered to euthanasia at week 1, 4, 8 and 16 respectively. The ultrastructural histopath ological changes in liver tissues taken from different sites were evaluated semiquan titatively. Ⅲ. One hundred and twenty seven times of 32 P GMS intrahepatic artery interventional therapies were performed on 93 patients with hepatic carcinoma, including 79 cases of primary hepatic carcinoma and 14 cases of secondary hepatic carcinoma. 32 P GMS ( n =30), and group B, 32 P GMS and half dose of trans hepatic artery embolization (TAE) ( n =49) , and 18 patients with HCC by TAE only as control group C. Fourteen patients with secondary hepatic carcinoma were treated in the same way as group B or C. RESULTS Ⅰ. Comparing with the control group, the treatment group of tumor bearing nude mice attained the tumor inhibition rates of 59 7%-93 7% ( F =579 62, P <0 01) at 14*!d . At an absorbed dose of 7320Gy, the tumor cells were completely destroyed. When the absorbed doses ranged from 1830Gy to 3660Gy, most of the tumor cells showed the evidences of injury or necrosis, but there appeared some well differentiated tumor cells and enhanced effect of the autoimmunocytes. At an absorbed dose of 366Gy or less, some tumor cells still remained active proliferative ability. The definite anticancer effect appeared as early as 3d after intratumoral injection of 32 P GMS. Ⅱ. The cumulative amount of 32 P GMS in the target tissue after trans hepatic artery instillation attained more than 90% of the total dose administrated. Semiquantitative analysis of ultrastructral morphology in the experimental group showed no statistical difference between the nuclear abnormality (N abn ) and mitochondrial variability (M var ) at week 1 or 2, but revealed prominent difference (χ 2=6 70-9 68, P <0 01 , χ 2=65 09-115 09, P <0 001 ) as compared with those in the other groups. In the experimental group the N abn in tissues showed no significant difference between week 8 and week 16. No apparent changes were found in the stomach, spleen, kidney and lung tissues of the experimental pigs. Ⅲ. The therapeutical results of HCC patients in group A were closely approximated to those of group C, no hematological toxic side effects were noted, and the systemic reaction was mild. In some patients 2*!mos - 3*!mos after treatment some secondary foci appeared around the periphery of the primary lesion. In general better effectiveness was obtained in patients with small lesion. After analyzing by RIDIT method, the therapeutic result in group B was significantly better than that in group C, and secondary foci around the original lesion were rarely seen at 3*!mos after treatment. In group C the collateral circulation was reestablished along the periphery of primary foci and the secondary foci appeared more frequently, and required to undergo several courses of treatment. In group B, 4 cases of HCC were treated surgically as their mass decreased in size after 32 P GMS treatment.

Segmental transcatheter arterial embolization for primary hepatocellular carcinoma

AIM To evaluate the therapeutic effects of segmental transcatheter arterial embolization for primary hepatocellular carcinoma, and to recognize the menifestation and clinical value of lipiodol overflow into portal veins surrounding the tumors. METHODS A total of 50 cases of nonresectable primary hepatocellular carcinoma underwent segmental transcatheter arterial embolization. Two methods of superselective segmental catheterization were used, one was the method of wire guiding, and the other the technic of co axial infusion catheter. RESULTS The 1 , 2 , 3 and 4 year cumulative survival rates of 50 cases with segmental transcatheter arterial embolization for primary hepatocellular carcinoma were 83 8%, 65 4%, 42 9% and 24 5% respectively. The incidence of the lipiodol overflow into portal veins was 64%. The overflow of lipiodol into portal veins, represented as 3-5 grade branches of portal veins visualized by lipiodol, was “star like” or “tree like”, and there was a relatively large vessel in the center surrounded with radicalized small branches of vessels. CONCLUSION The lipiodol overflow into portal veins was one of the signs of complete embolization for tumors, and may play a partial role in embolizating the portal venous supply for hepatocellular carcinoma.

Some recent works on diagnosis and treatment of gastric cancer

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Portal vein embolization by fine needle ethanol injection :experimental and clinicalstudies

AIM To improve the technique of intraportal embolization (PVE) therapy, a new embolic method, was devised and the safety, effectiveness and feasibility were evaluated. METHODS PVE with intraportal ethanol injection via a fine needle was performed in 28 normal dogs, 22 SD rats, and 24 cirrhotic SD rats. After PVE, portography, histological and functional alteration of the liver were evaluated in dogs and rats, and the changes in portal hemodynamics as well as hepatic anatomy were observed in rats. In the clinical study, PVE by ethanol injection was performed in 61 patients with hepatocellular carcinoma under the guidance of portoechography with intraportal injection of CO 2. The effect of PVE was evaluated by ultrasonography and laparotomy. RESULTS The effectiveness and toxicity were dependent on the dose of ethanol. In the dogs, 0 25*!mg/*!kg of ethanol caused incomplete embolization with least liver damage, while 1 0*!mg/*!kg induced complete embolization with a high mortality of 57 1% (4/*!7) due to respiratory arrest. The dose of 0 5*!mg/*!kg resulted in complete embolization with slight toxicity to the liver. In the rats, the survival rate was 100% in normal group but 40 9% in cirrhotic models after ethanol injection by dose of 0 05*!mg/*!100*!g . PVE for cirrhotic rats with 0 03*!mg/*!100*!g of ethanol induced satisfactory embolization with significant hypertrophy in nonembolized lobes, and only slight damage to the hepatic parenchyma, and transient alteration in liver function, portal pressure and portal flow. In the clinical study, 12 cases with reverse portal flow were excluded judged by portoechography. Satisfactory embolization was gained in 90 2% (55/*!61) of the remaining patients determined by ultrasonography and surgery. All cases ran an uneventful postembolization course with no aberrant embolization. CONCLUSION PVE with intraportal ethanol injection of appropriate dosage via a fine needle is safe and effective and has several advantages comparing with transcatheter method. Portoechography is a mandatory approach for the prevention of aberrant embolization.

The therapeutic effects of recombinant adenovirus RA538 on human gastric carcinoma cells in vitro and in vivo

AIM To evaluate the potential of RA-538 genetherapy for gastric carcinoma.METHODS Human gastric carcinoma cell lineSGC7901 treated with Ad-RA538 or Ad-LacZ wereanalysed by X-gal stain,MTT,DNA ladder,Tunel,flow cytometric analysis,PCR,andWestern Blot in vitro.The tumorigenicity andexperimental therapy in nude mice model wereassessed in vivo.RESULTS Ad-LacZ could efficiently transferthe LacZ gene into SGC7901 cells.X-gal-positivecells at MOI 25,50,100,and 200 were 90%,100%,100%,and 100% respectively.Ad-RA538could strongly inhibit cell growth and inducedapoptosis in SGC7901 cells.The proliferation ofthe Ad-RA538-infected SGC7901 cells wasreduced by 76.3%.The mechanism of killing ofgastric carcinoma cells by Ad-RA538 was foundto be apoptosis by DNA ladder,Tunel and flowcytometric analysis.The tumorigenicity in nudemice using Ad-RA538 showed that all three micefailed to form tumor from 7 to 30 days comparedwith Ad-LacZ and parent SGC7901 cells.Experimental therapy on the nude mice modelbearing subcutaneous tumor of SGC790| cells showed that intratumor instillation of Ad-RA538inhibited the growth of the tumors.Ad-RA538-treated tumors were inhibited by 60.66 %,compared with that of the tumor injected withAd-LacZ and mock.CONCLUSION The expression of Ad-RA538 can inhibit growth and induce apoptosis of gastric cancer cell in vitro and in vivo. Ad-RA538 can be used potentially in gene therapy for gastric carcinoma.

Clinical research advances in primary liver cancer

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Progress in research of liver surgery in China

INTRODUCTIONLiver surgery,was started in the late 1950s in Chinaand has developed rapidly in the past 40 years.The study on the diagnosis and treatment of primaryliver cancer in China underwent four stages:①Inthe 1950s,the anatomical study of the liver lay asolid foundation for liver resection.①In

Gastroesophageal reflux and Helicobacter pylori:a review

INTRODUCTION Since the observation by Labenz et al thateradication of Helicobacter pylori(Hp)infectionmay be followed by development of refluxesophagitis in a relevant proportion of duodenalulcer patients previously not affected bygastroesophageal reflux disease(GERD),agrowing attention has been given to the

Expression of bcl-2 oncogene in gastric precancerous lesions and its correlation with syndromes in traditional Chinese medicine

AIM: To observe the protein and mRNA expression of bcl-2 oncogene in gastric precancerous lesions (GPL) and to analyze its correlation with syndromes in traditional Chinese medicine (TCM). METHODS: Sixty-seven patients with GPL confirmed by gastroscopy and pathology were studied,including 39 cases of moderate gastric mucosal dysplasia,19 cases of severe gastric mucosa dysplasia, 9 cases of incomplete colon metaplasia. In syndrome differentiation of TCM,17 cases belonged to the syndrome of qi and yin deficiency of the spleen and stomach complicated by qi stagnation, 21 cases belonged to the syndrome of qi and yin deficiency of the spleen and stomach complicated by stomach heat, 29 cases belonged to the syndrome of qi and yin deficiency of the spleen and stomach complicated by blood stasis. Protein and mRNA expression of bcl-2 oncogene were detected by labeled streptavidin biotin (LSAB) immunohist-ochemistry and in situ hybridization respectively. RESULTS: Abnormal expression of protein and mRNA on bcl-2 oncogene was found in GPL,which increased gradually with the course of lesions. In moderate and severe gastric mucosal dysplasia and incomplete colon metaplasia,there was no difference in the expression of bcl-2 oncogene (P>0.05).In different accompanying syndromes, the expression of protein and mRNA on bcl-2 oncogene increased gradually in the following order: deficiency of both qi and yin of the spleen and stomach accompanying qi stagnation→stomach heat→blood stasis. In GPL, compared with accompanying blood stasis, there was an obvious difference in the expression of bcl-2 oncogene between the syndrome of qi and yin deficiency of the spleen and stomach and accompanying stomach heat, so did accompanying qi stagnation (the level of protein: X2=8.45, P<0.05;the level of mRNA: X2=7.35, P<0.05). CONCLUSION: Apoptosis-associated bcl-2 oncogene is abnormally expressed in GPL,which correlates with different accompanying syndromes in TCM.

Source of blood supply of liver cavernous hemangioma and sclerosis and embolization treatment

AIM To investigate the source of blood supply of carvenous hemangioma of liver (CHL) and provide a feasibile treatment for CHL via hepatic artery. METHODS Ⅰ. Origin of blood supply of CHL: portovenography, hepatic arteriography and portal vein staining were performed in 5 patients. Two casts of hepatic blood vessels from resected specimen were observed. Ⅱ. Clinical data: Among 75 patients (30 males, 45 females, aged 25～57 years with a mean of 37 4). 56 were of solitary type (44 on the right lobe, 12 on the left with 4 having intraparenchymatoma) and 19 were of multiple type (9 on the right, 2 the left, 8 whole liver). Twenty two patients were treated by sclerosis, 50 by embolization via hepatic artery and 3 were excised. RESULTS In 5 cases with portography, the contrast medium did not enter the tumor, the tumor appeared as low denty area and the intrahepatic branches of portal vein were pushed aside. In 5 cases with portal vein staining, the normal liver parenchyma was stained deep blue, and the tumor was not stained. The tumor area appeared as a round vacant cavity in 2 specimen casts. In 72 patients treated with sclerosis a or embolization via hepatic artery or through interventional method, the tumors diminished by 10%～30% in diameter and no tumors grew larger. CONCLUSION The blood supply of CHL originates from the hepatic artery. Tumors treated with sclerosis and emblization decreased in size or got fiberized.

The effect pathway of retinoic acid through regulation of retinoic acid receptor α in gastric cancer cells

AIM To evaluate the role of RARα gone in mediating the growth inhibitory effect of all-trans retinoic acid(ATRA) on gastric cancer cells. METHODS The expression levels of retinoic acid receptors(PARs)in gastric cancer cells were detected by Northern blot.Transient transfection and chlorophenicol acetyl transferase(CAT)assay were used to show the transcriptional activity of β retinoic acid response element (βPARE)and AP-1 activity.Cell growth inhibition was determined by MTT assay and anchorage-independent growth assay,respectively.Stable transfection was performed by the method of Lipofectamine,and the cells were screened by G418. RESULTS ATRA could induce expression level of RARα in MGC80-3,BGC-823 and SGC-7901 cells obviously, resulting in growth inhibition of these cell lines.After sense RARα gone was transfected into MKN-45 cells that expressed rather low level of RARα and could not be induced by ATPA,the cell growth was inhibited by ATPA markedly.In contrast,when antisense RARα gone was transfected into BGC-823 cells,a little inhibitory effect by ATPA was seen,compared with the parallel BGC-823 cells.In transient transfection assay,ATPA effectively induced transcriptional activity of βRARE in MGC80-3, BGC-823,SGC-7902 and MKN/RARα cell lines,but not in MKN-45 and BGC/aRARα cell lines.Similar results were observed in measuring anti-AP-1 activity by ATPA in these cancer cell lines. CONCLUSION ATRA inhibits the growth of gastric cancer cells by up-regulating the level of RARα; RARα is the major mediator of ATRA action in gastric cancer cells;and adequate level of RARα is required for ATRA effect on gastric cancer cells.